Heterocyclic derivatives and their use as antithrombotic agents

ABSTRACT

The present invention relates to antithrombotic compounds comprising the group Q, Q having formula (I), wherein the substructure (i) is a structure selected from (a, b and c), wherein X is 0 or S; X′ being independently CH or N; and m is 0, 1, 2 or 3; wherein the group Q is bound through an oxygen atom or an optionally substituted nitrogen or carbon atom, or a pharmaceutically acceptable salt thereof or a prodrug thereof. The compounds of the invention are therapeutically active and in particular are antithrombotic agents.

[0001] The invention relates to new therapeutic compounds, in particularto antithrombotic agents, a process for their preparation,pharmaceutical compositions containing the compounds as activeingredients, as well as the use of said compounds for the manufacture ofmedicaments.

[0002] In therapy, a multiplicity of active compounds is used for thetreatment and prophylaxis of all sorts of diseases. Drugs differ widelyin their pharmacodynamic effects and clinical application, inpenetrance, absorption and usual route of administration, indistribution among the body tissues and in disposition and mode ofaction.

[0003] Apart from the type of patient and the type of disease to betreated or to be prevented, the physicochemical properties oftherapeutically active compounds determine to a great extent thepreferred route of their admistration. In the development of drugs, theoral applicability thereof is usually an important selection criterium.For the majority of patients this is obviously the most convenient routefor access of the drug to the systemic circulation. In order for adrug—administered via oral route—to act, it must first be absorbedbefore it is transported to the appropriate tissue or organ, where itmay penetrate to the responding subcellular structure and maysubsequently be metabolized, or where it may be bound, stored, orwhatever is necessary to elicit a response or to alter ongoingprocesses. However, not always compounds which have been found topossess an advantageous therapeutic activity are also sufficientlyabsorbed in the gastrointestinal tract to display effective oralbioavailability.

[0004] Thus, one of the pivotal issues in drug design is to developcompounds which both show activity and good absorptive properties. Animportant area in which is actively sought for oral biavailability isthe area of antithrombotic agents.

[0005] The present invention relates to antithrombotic compoundscomprising the group Q, Q having the formula

[0006] wherein the substructure

[0007] is a structure selected from

[0008] wherein X is O or S;

[0009] X′ being independently CH or N;

[0010] and m is 0, 1, 2 or 3;

[0011] wherein the group Q is bound through an oxygen atom or anoptionally substituted nitrogen or carbon atom,

[0012] or a pharmaceutically acceptable salt thereof or a prodrugthereof

[0013] The compounds of the invention are active antithrombotic agentshaving an improved pharmacological profile, in particular with regard toproperties like their absorptive properties and their toxicity.

[0014] The term “antithrombotic compound” means any compound havingantithrombotic activity. Examples of such compounds are inhibitors ofserine proteases that play a role in the blood coagulation cascade orGpIIb/IIIa antagonists.

[0015] The group Q is bound to the molecule through an oxygen atom or anoptionally substituted nitrogen or carbon atom. “Optionally substituted”in this respect means any suitable substituent, such as, but not limitedto, oxo, alkyl, alkenyl, alkoxy, aryl, halogen and the like,

[0016] The term “prodrug” means a compound of the invention in which(an) amino group(s) is (are) protected, e.g. by (a) hydroxy or(1-6C)alkoxycarbonyl group(s), or a compound wherein—if present— (a)carboxylate group(s) is (are) esterified.

[0017] The present invention relates to the surprising finding that thepresence of the group Q in antithrombotic compounds gives rise tofavourable properties of the compounds. Especially when Q is used toreplace a basic moiety in compounds of which is known that they requiresuch a moiety for their activity, an improvement of the pharmacologicalproperties is realized, and in particular when that basic moiety is a(hetero)arylguanidino or (hetero)arylamidino moiety. In particular animprovement of the absorptive properties of those compounds is observed.Also a reduction of the toxicity of compounds of this invention isobserved.

[0018] Preferably the group Q has the formula

[0019]

[0020] m being as previously defined.

[0021] An established in vitro model for the determination of theabsorptive properties of drugs is the Caco-2 model (Artursson, P.,S.T.P. Pharma Sciences 3(1), 5-10, 1993; Walter, E. et al.Pharmaceutical Research, 3, 360-365, 1995). In this in vitro model thetransepithelial transport properties of a compound are determined inmonolayers of a human intestinal cell-line (Caco-2) in terms of apermeability coefficient (apparent permeability). This model is usefulfor the prediction of in vivo absorption of compounds in thegastrointestinal tract. Preferably the antithrombotic compounds of theinvention have a Caco-2 permeability of 8 nm/s or higher.

[0022] As noted above, amongst the compounds of the present inventionare inhibitors of serine proteases of the blood coagulation cascade, andin particular inhibitors of thrombin and/or factor Xa. Preferredcompounds inhibit thrombin more effectively than other serine proteases.More preferred compounds are thrombin inhibitors having, in addition, anIC₅₀ value of less than 1 μM. The compounds are useful for treating andpreventing thrombin-mediated and thrombin-associated diseases. Thisincludes a number of thrombotic and prothrombotic states in which thecoagulation cascade is activated which include, but are not limited to,deep vein thrombosis, pulmonary embolism, thrombophlebitis, arterialocclusion from thrombosis or embolism, arterial reocclusion during orafter angioplasty or thrombolysis, restenosis following arterial injuryor invasive cardiological procedures, postoperative venous thrombosis orembolism, acute or chronic atherosclerosis, stroke, myocardialinfarction, cancer and metastasis, and neurodegenerative diseases.Compounds of the invention may also be used as ill vitro anticoagulantsor as anticoagulants in extracorporeal blood circuits, such as thosenecessary in dialysis and surgery.

[0023] Serine proteases are enzymes which play an important role in theblood coagulation cascade. Apart from thrombin and factor Xa, otherexamples of this group of proteases comprise the factors VIIa, IXa, XIa,XIIa, and protein C.

[0024] Thrombin is the final serine protease enzyme in the coagulationcascade. The prime function of a thrombin is the cleavage of fibrinogento generate fibrin monomers, which are cross-linked to form an insolublegel. In addition, thrombin regulates its own production by activation offactors V and VIII earlier in the cascade. It also has important actionsat cellular level, where it acts on specific receptors to cause plateletaggregation, endothelial cell activation and fibroblast proliferation.Thus thrombin has a central regulatory role in homeostasis and thrombusformation. Since inhibitors of thrombin may have a wide range oftherapeutical applications, extensive research is done in this area.

[0025] In the development of synthetic inhibitors of serine proteases,and more specifically of thrombin, the benzamidine moiety is one of thekey structures. It mimics the protonated side-chain of the basic aminoacids Arg and Lys of its natural substrates. Compounds with this moietyhave been studied extensively and repeatedly. A very potentrepresentative of this type of thrombin inhibitors is the amino acidderivativeNα-(2-naphthylsulfonyl)-glycyl-4-amidinophenylalanin-piperidide (NAPAP)(Stürzebecher, J. et al., Thromb. Res. 29, 635-642, 1983). However, thepharmacological profile of NAPAP is unattractive for therapeuticalapplications: the compound shows toxic effects after intravenousadministration and, in addition, poor oral bioavailability after oraladministration. Up until now, the NAPAP-like benzamidine derivativeswhich have been investigated for use as thrombin inhibitors show theseunfavourable pharmacological and pharmacokinetic properties. It wasassumed that these properties are due to the strong basicity of theamidino functionality of these compounds (Kaiser, et al., Pharmazie 42,119-121, 1987; Stürzebecher, J. et al., Biol. Chem. Hoppe-Seyler, 373,491-496, 1992). Several studies have been performed on variations ofthis basic structure (see for example Stürzebecher, J. et al., Pharmazie43, 782-783, 1988; Stürzebecher, J. et al. (1993), DIC-Pathogenesis,Diagnosis and Therapy of Disseminated Intravascular Fibrin Formation [G.Müller-Berghaus et al., eds.] pp. 183-190, Amsterdam, London, New York,Tokyo: Exerpta Medica). However, modifications of the benzamidine moietydecreasing its basicity always resulted in a drastic loss ofanti-thrombin activity (Stürzebecher, J. et al., J. Enzyme Inhibition 9,87-99, 1995).

[0026] Oral bioavailability is a property of thrombin inhibitors whichis urgently searched for. Potent intravenous thrombin inhibitors areclinically effective in acute care settings requiring the treatment ofthrombin-related diseases However, particularly the prevention ofthrombin-related diseases such as myocardial infarct, thrombosis andstroke require long-term therapy, preferably by orally dosing ananticoagulant. Consequently, the search for active, orally bioavailablethrombin inhibitors continues unabated. Oral bioavailability is at leastin part related to the ability of compounds to be absorbed in thegastrointestinal tract. The low oral bioavailability of NAPAP and itsanalogues may therefore be related to their deficient absorptiveproperties in the intestines.

[0027] The present invention provides a solution to the deficientpharmacological properties of the NAPAP-like compounds, in particularwith respect to the toxicity and the deficient absorptive properties.

[0028] Preferred serine protease inhibitors of the invention have theformula (I), comprising the group Q; compounds of this type showimproved transepithelial transport properties (increased apparentpermeability) in comparison with prior art compounds:

R¹—Y—[NR²—A—C(O)]_(n)—NR³—CHR⁴—C(O)—R⁵   (I)

[0029] wherein R¹ is (1-8C)alkyl, (6-14C)aryl-(1-8C)alkenyl,(6-14C)aryl-(1-8C)alkanoyl, (6-14C)aryl, (7-15C)aralkyl, bisaryl,heteroaryl, heteroaralkyl(1-8C)alkyl, heterocycloalkyl, cycloalkyl orcycloalkyl substituted alkyl; R² is H or (1-8C)alkyl; R³ is Q when R⁴ isH, or R³ is H or (1-8C)alkyl when R⁴ is Q; Q is as previously defined;R⁵ is OH or OR⁶, R⁶ being (1-8C)alkyl (3-12C)cycloalkyl or(7-15C)aralkyl, or R⁵ is NR⁷R⁸, wherein R⁷ and R⁸ are the same ordifferent being H, (1-8C)alkyl, (3-12C)cycloalkyl, (6-14C)aryl,(7-15C)aralkyl, optionally subtituted with (1-8C)alkoxy, C(O)OH orC(O)OR⁶, or R⁷ and R⁸ together with the nitrogen atom to which they arebonded are a nonaromatic (4-8)membered ring optionally containinganother heteroatom, which ring may be condensed with another optionallyaromatic ring and may be subtituted with OH, an oxo group, (1-8C)alkyl,optionally substituted with one or more halogens or hydroxy groups,(2-8C)alkenyl, (1-8C)alkylidene, (2-8C)alkynyl, (1-8C)alkoxy,(1-8C)acyl, (6-14C)aryl, C(O)OH, C(O)OR⁶, C(O)NR⁹R¹⁰, wherein R⁹ and R¹⁰are the same or different being H or (1-8C)alkyl, or SO₂R¹¹ and R¹¹ is(1-8C)alkyl optionally substituted by one or more fluorine atoms; Y isSO₂ or C(O); A is CHR¹², R¹² being H, phenyl, benzyl, (1-8C)alkyl,optionally substituted with OH or COR¹³ wherein R¹³ is OH, (1-8C)alkoxy,morpholino, morpholino(1-8C)alkoxy, NH₂, NHR¹⁴ or NR¹⁴R¹⁵, R¹⁴ and R¹⁵being independently (1-8C)alkyl optionally substituted with C(O)OR² orR¹⁴ and R¹⁵ are a nonaromatic (4-8)membered ring together with thenitrogen atom to which they are bonded, or R¹² together with R³ is—(CH₂)_(s)— when R⁴ is Q, s being 2, 3 or 4, or A is NR²;

[0030] and n is 0 or 1, or a pharmaceutically acceptable salt thereof.

[0031] Related thrombin inhibitors are disclosed in WO 92/16549 and WO92/08709, wherein respectively para- and meta-substituted phenylalaninederivatives are described having an amidino, guanidino, oxamidino,aminomethyl or amino substituent. However, compounds with the amidinosubstituent show unfavourable pharmacological properties, whereas theother structures, with a modified amidino moiety, display a loss ofactivity (vide sipra). Other modifications are described in EP 555824where compounds are disclosed having a benzimidazolyl group, whichcompounds do not contain a primary amino functionality.

[0032] Thrombin inhibitors having a benzamidine moiety have also beenmodified in other parts of the molecule, however, without improvement ofthe unfavourable pharmacological properties caused by the amidinosubstituent. Examples hereof are disclosed in EP 508220, wherein thecompounds contain an azaglycyl group instead of the glycyl group ofNAPAP; in DE 4115468, wherein that glycyl group is replaced e.g. by anaspartyl group; in WO 94/18185, wherein no glycyl group is present inthe compounds and the piperidine group which is present in NAPAP isreplaced by a piperazide group; in WO 95/13274, wherein the compoundsalso do not have a glycyl group and modifications are made—in comparisonwith NAPAP—in the arylsulfonyl part and the piperidine part of themolecule, and in EP 236163, wherein Nα-alkyl substitutedamidinophenylalanin derivatives are described. Furthermore,Stürzebecher, J. et al. (Thrombosis Research 54, 245-252, 1989) suggestthat the alkylene linkage connecting the benzamidine moiety to the restof the molecule may have a length of 1-3 methylene groups.

[0033] Therefore, there is a still need for serine protease inhibitors,especially thrombin inhibitors, with more favourable pharmacologicalproperties, such as the inhibitors of the present invention whichpotentially have good oral bioavailability

[0034] Preferred compounds of formula (I) are compounds wherein R¹ isphenyl, naphthyl, (iso)quinolinyl, tetrahydro(iso)quinolinyl,3,4-dihydro-1H-isoquinolinyl, 2,3,4,5-tetrahydro-1H-benzo[d]azepinyl,2,3-dihydro-5H-benzo[f][1,4]oxazepinyl, dibenzofuranyl, chromanyl,bisaryl, each aryl being a 5- or 6-membered ring and optionallycontaining a O, S or N-atom, which groups R¹ may optionally besubstituted with one or more substituents selected from (1-8C)alkyl,(1-8C)alkoxy the alkyl group of which may be optionally substituted withan alkoxy group or an alkoxyalkyl group, phenyl-(1-8C)alkyl,tetrahydropyranyloxy, tetrahydropyranyloxy(1-8C)alkyl or NR¹⁵R¹⁶, inwhich R¹⁵ and R¹⁶ are independently selected from H and (1-8C)alkyl, orR¹ is (1-8C)alkyl substituted with a (5-8C)cycloalkyl,(7-10C)bicycloalkyl or (10-16C)polycycloalkyl, optionally substitutedwith a group selected from oxo or (1-8C)alkyl; R³ is H or (1-8C)alkyl;R⁴ is Q; R⁵ is (1-8C)alkoxy, NR⁷R⁸, wherein R⁷ and R⁸ are the same ordifferent being H, (1-8C)alkyl, (3-12C)cycloalkyl, optionallysubstituted with (1-8C)alkoxy or COOR⁶, or R⁵ is a group of the formula

[0035] wherein the interrupted line represents an optional bond, B beingCR¹⁷ when the optional bond is present, or B is CHR¹⁷, R¹⁷ being H,(1-8C)alkyl, optionally substituted with one or more halogens or hydroxygroups, (2-8C)alkenyl, (2-8C)alkynyl, (1-8C)acyl, or (1-8C)alkoxy, or Bis O, S, or NR¹⁸, R¹⁸ being (1-8C)alkyl, (1-8C)acyl, C(O)NR⁹R¹⁰ orSO₂-(1-8C)alkyl optionally substituted by one or more fluorine atoms.

[0036] More preferred compounds of formula (I) are compounds wherein R²and R³ are H and Q is

[0037] Also preferred are compounds wherein R⁵ is

[0038] and B is CH₂ or CH(1-8C)alkyl.

[0039] Preferred groups R¹ are phenyl, naphthyl,tetrahydroisoquinolinyl, 3,4-dihydro-1H-isoquinolinyl,2,3,4,5-tetrahydro-1H-benzo[d]azepinyl,2,3-dihydro-5H-benzo[f][1,4]oxazepinyl, which groups R¹ may optionallybe substituted with one or more substituents selected from (1-8C)alkyl,(1-8C)alkoxy the alkyl group of which may be optionally substituted withan alkoxy group or an alkoxyalkyl group, phenyl-(1-8C)alkyl,tetrahydropyranyloxy, tetrahydropyranyloxy(1-8C)alkyl or NR¹⁵R¹⁶. Incompounds with these preferred R¹ groups, Y is preferably SO₂.

[0040] When R⁴ is

[0041] n preferably has the value 1. Preferably A is CH(1-8C)alkylsubstituted with COR¹³ wherein R¹³ is OH, (1-8C)alkoxy, morpholino,morpholino(1-8C)alkoxy, NHR¹⁴ or NR¹⁴R¹⁵, R¹⁴ and R¹⁵ beingindependently (1-8C)alkyl, or A is CHR¹², R¹² being —(CH₂)_(s)— togetherwith R³ when R⁴ is Q, s being 2 or 3.

[0042] Other preferred compounds are compounds wherein n is 0 and R⁴ is

[0043] X being as previously defined.

[0044] Most preferred are the compounds according to formula (I) whereinR¹ is

[0045] n is 1, R² is H, A is CHCH₂C(O)OH, CHCH₂C(O)O(1-8C)alkyl,CHCH₂C(O)morpholine, CHCH₂C(O)O(1-8C)alkylene-morpholine, CHCH₂C(O)NHR¹⁴or CHCH₂C(O)NR¹⁴R¹⁵, R¹⁴ and R¹⁵ being independently (1-8C)alkyl, R³ isH, R⁴ is

[0046] and R⁵ is

[0047] or the compounds according to formula (I) wherein R¹ is selectedfrom

[0048] In the description of the compounds of formula (I) the followingdefinitions are used. The term (1-8)alkyl means a branched or unbranchedalkyl group having 1-8 carbon atoms, for example methyl, ethyl, propyl,isopropyl, butyl, sec-butyl, tert-butyl, hexyl and octyl. The term(2-8C)alkenyl means a branched or unbranched alkenyl group having 2 to 8carbon atoms, such as ethenyl, 2-butenyl, etc.. The term (1-8C)alkylenemeans a branched or unbranched alkylene group having 1 to 8 carbonatoms, such as —(CH₂)_(α)— wherein α is 1 to 8, —CH(CH₃)—,—CH(CH₃)—CH₂—, etc.. The term (1-8C)alkylidene means a branched orunbranched alkylidene group having 1-8 carbon atoms, such as methyleneand ethylidene. The term (2-8C)alkynyl means a branched or unbranchedalkynyl group having 2-8 carbon atoms, such as ethynyl and propynyl. Theterm (3-12C)cycloalkyl means a mono- or bicycloalkyl group having 3-12carbon atoms, being cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, cyclo-octyl, the camphor group, etc.. A preferredcycloalkyl group in the definition of R¹ is the camphor group. The term(1-8C)alkoxy means an alkoxy group having 1-8 carbon atoms, the alkylmoiety having the meaning as previously defined. The term (1-8C)acylmeans an acyl group having 1-8 carbon atoms, the alkyl moiety having themeaning as previously defined. Formyl and acetyl are preferred acylgroups. The term (1-8)alkanoyl means an oxo-alkyl group, the alkylmoiety having the meaning as previously defined.

[0049] The term (6-14C)aryl means an aromatic hydrocarbon group having 6to 14 carbon atoms, such as phenyl, naphthyl, tetrahydronaphthyl,indenyl, which may optinally be substituted with one or moresubstituents such as -but not limited to- alkyl, alkoxy, the alkyl groupof which may be optionally substituted with an alkoxy group or analkoxyalkyl group (e.g the substituent groups —O—(CH₂)₂—OCH₃ or—O—CH(CH₂OCH₃)₂), tetrahydropyranyloxy, tetrahydropyranyloxymethyl,acyl, alkylthio, hydroxyalkyl, haloalkyl, carboxy, carboxyalkyl,carboalkoxy, hydroxy, halogen, trifluoromethyl, trifluoromethylcarbonyl,nitro, cyano, amino, dialkylamino, alkylsulfinyl and/or alkylsulfonyl(in the relevant cases alkyl is meant to be (1-8C)alkyl).

[0050] Preferred aryl groups are

[0051] The term (7-15C)aralkyl means an aralkyl group having 7 to 15carbon atoms, wherein the alkyl group is a (1-8C)alkyl group and thearyl group is a (6-14C)aryl as previously defined. Phenyl-(1-8C)alkylgroups are preferred aralkyl groups, such as benzyl.

[0052] The term heteroaryl means a substituted or unsubstituted aromaticgroup having 4 to 12 carbon atoms, at least including one heteroatomselected from N, O, and S, like imidazolyl, thienyl, benzthienyl,quinolinyl, isoquinolinyl, tetrahydroquinolinyl,tetrahydroisoquinolinyl, indolyl, dibenzofuranyl, chromanyl,

[0053] The substituents on the heteroaryl group may be selected from thegroup of substituents listed for the aryl group.

[0054] Heteroaralkyl groups are analogs of the (7-15C)aralkyl groups, atleast including one heteroatom selected from N, O, and S

[0055] The term bisaryl in the definition of R¹ means two independentlychosen aryl or heteroaryl groups according to the definitions of theterm aryl and heteroaryl, connected to each other by a bond or by ashort bridge, having a length of one or two atoms, such as CH₂, N₂ orSO₂, optionally substituted with a substituent as listed for the arylgroup. Examples of bisaryls are biphenyl,

[0056] The term cycloalkyl substituted alkyl in the definition of R¹means an alkyl group, preferably having 1-8 carbon atoms, carrying amono-, bi- or polycycloalkyl group, preferably having 5-8, 7-10 and10-16 carbon atoms, respectively, which cycloalkyl group may optionallybe substituted with an oxo group and/or a substituent as listed for thearyl group.

[0057] The term heterocycloalkyl means an optionally substitutedcycloalkyl group, preferably having 4 or 5 carbon atoms, furthercontaining one heteroatom selected from O, S or N, such astetrahydrofuran and tetrahydropyran. The substituents on thehetercycloalkyl group may be selected from the group of substituentslisted for the aryl group.

[0058] The term nonaromatic (4-8)membered ring in the definition ofNR⁷R⁸, where R⁷ and R⁸ together with the nitrogen atom to which they arebonded are a ring, means a ring containing the nitrogen atom and furtherhaving at most 3-7 carbon atoms, which ring may contain unsaturatedbonds. Examples of such (4-8)membered rings are azetidine, pyrrolidine,piperidine, piperazine morpholine and thiomorpholine.

[0059] In the development of synthetic inhibitors of serine proteases,and more specifically of thrombin, the interest in small syntheticpeptides that are recognized by proteolytic enzymes in a manner similarto that of natural substrates, has increased. As a result, newpeptide-like inhibitors have been prepared, such as the transition stateinhibitors of thrombin and the low molecular weight thrombin inhibitorInogatran (Thromb. Haemostas. 1995, 73:1325 (Abs. 1633); WO 93/11152(Example 67)), which has been disclosed to be a potent and selectivethrombin inhibitor. Related compounds are described in WO 95/23609; incomparison with Inogatran and its analogs, compounds disclosed in thispatent application have an aromatic group in the agmatine-like group.Although these developments already have lead to new and meaningfulinsights, the search for more effective and more selective, and inparticular orally applicable, thrombin inhibitors still proceeds.

[0060] Thus, other preferred serine protease inhibitors of the inventionare small synthetic peptides comprising the group Q, and have theformula (XX)

J—D—E—N(R′)¹—Q   (XX)

[0061] in which formula Q is as previously defined; (R′)¹ is H or(1-4C)alkyl; J is H, optionally substituted D,L α-hydroxyacetyl, (R′)²,(R′)²—O—C(O)—, (R′)²—C(O)—, (R′)²—SO₂—, (R′)⁷OOC—(CH(R′)³)_(p)—SO₂—,(R′)⁷OOC—(CH(R′)³)_(p)—, (R′)³ ₂NCO—(CH(′)³)_(p)—,Het-CO—(CH(R′)³)_(p)—, or an N-protecting group, wherein (R′)² isselected from (1-12C)alkyl, (2-12C)alkenyl, (2-12C)alkynyl and(3-12C)cycloalkyl, which groups may optionally be substituted with(3-12C)cycloalkyl, (1-6C)alkoxy, oxo, OH, COOH, CF₃ or halogen, and from(6-14C)aryl, (7-15C)aralkyl and (8-16)aralkenyl, the aryl groups ofwhich may optionally be substituted with (1-6C)alkyl, (3-12C)cycloalkyl,(1-6C)alkoxy, OH, COOH, CF₃ or halogen; each group (R′)³ isindependently H or has the same meaning as (R′)²; (R′)⁷ has the samemeaning as (R′)³ or is Het-(1-6C)alkyl or Het-(2-6C)alkynyl; and Het isa 4-, 5- or 6-membered heterocycle containing one or more heteroatomsselected from O, N or S; p is 1, 2 or 3; D is a bond, an amino-acid ofthe formula —NH—CH[(CH₂)_(q)C(O)OH]—C(O)— or an ester derivative thereofand q being 0, 1, 2 or 3, —N((1-12C)alkyl)—CH₂—CO—,—N((2-12C)alkenyl)—CH₂—CO—, —N((2-12C)alkynyl)—CH₂—CO—,—N(benzyl)—CH₂—CO—, D-1-Tiq, D-3-Tiq, D-Atc, Aic, D-1-Piq, D-3-Piq or aL- or D-amino acid having a hydrophobic, basic or neutral side chain,which amino acid may optionally be N—(1-6C)alkyl substituted; or J and Dtogether are the residue (R′)⁴(R′)⁵N—CH(R′)⁶—C(O)—, wherein (R′)⁴ and(R′)⁵ independently are (R′)², (R′)²—O—C(O)—, (R′)²—C(O)—, (R′)²—SO₂—,(R′)⁷OOC—(CH(R′)³)_(p)—SO₂—, (R′)⁷OOC—(CH(R′)³)_(p)—,H₂NCO—(CH(R′)³)_(p)—, or an N-protecting group, or one of (R′)⁴ and(R′)⁵ is connected with (R′)⁶ to form a 5- or 6-membered ring togetherwith “N—C” to which they are bound, which ring may be fised with analiphatic or aromatic 6-membered ring; and (R′)⁶ is a hydrophobic, basicor neutral side chain; E is an L-amino acid with a hydrophobic sidechain, serine, threonine, a cyclic amino acid optionally containing anadditional heteroatom selected from N, O or S, and optionallysubstituted with (1-6C)alkyl, (1-6C)alkoxy, benzyloxy or oxo, or E is—N(R′)³—CH₂—C(O)— or the fragment

[0062] wherein t is 2, 3, or 4, and W is CH or N, or E—N(R′)¹ takentogether form the fragment

[0063] wherein u is 1, 2 or 3; or a prodrug thereof

[0064] Preferred compounds of formula (XX) are those wherein E is anL-amino acid with a hydrophobic side chain, serine, threonine or—N(R′)³—CH₂—C(O)— or wherein E—N(R′)¹ taken together form the fragment

[0065] Other preferred compounds have the formula (XX), wherein J is aspreviously defined; D is a bond, an amino-acid of the formula—NH—CH[(CH₂)_(q)C(O)OH]—C(O)— or an ester derivative thereof and q being0, 1, 2 or 3, —N((1-6C)alkyl)—CH₂—CO—, —N((2-6C)alkenyl)—CH₂—CO—,—N(benzyl)—CH₂—CO—, D-1-Tiq, D-3-Tiq, D-Atc, Aic, D-1-Piq, D-3-Piq or aD-amino acid having a hydrophobic side chain, which amino acid mayoptionally be N-(1-6C)alkyl substituted;

[0066] or J and D together are the residue (R′)⁴(R′)⁵N—CH(R′)⁶—C(O)—;and E is a cyclic amino acid optionally containing an additionalheteroatom selected from N, O or S, and optionally substituted with(1-6C)alkyl, (1-6C)alkoxy, benzyloxy or oxo, or E is —N(R′)³—CH₂—C(O)—or the fragment

[0067] More preferred compounds of formula (XX) are those wherein J isH, 2-hydroxy-3-cyclohexyl-propionyl-, 9-hydroxy-fluorene-9-carboxyl,(R′)², (R′)²—SO₂—, (R′)⁷OOC—(CH(R′)³)_(p)—SO₂—, (R′)⁷OOC—(CH(R′)³)_(p)—,(R′)³ ₂N—CO—(CH(R′)³)_(p)—, Het-CO—(CH(R′)³)_(p)— wherein Het containsas a heteroatom at least a nitrogen atom which is bound to CO, or anN-protecting group, wherein (R′)² is selected from (1-12C)alkyl,(2-12C)alkenyl, (6-14C)aryl, (7-15C)aralkyl and (8-16)aralkenyl whichgroups may optionally be substituted (1-6C)alkoxy; each group (R′)³ isindependently H or has the same meaning as (R′)²; (R′)⁷ has,the samemeaning as (R′)³ or is morpholino-(1-6C)alkyl ormorpholino-(2-6C)alkynyl; D is a bond, D-1-Tiq, D-3-Tiq, D-Atc, Aic,D-1-Piq, D-3-Piq or a D-amino acid having a hydrophobic side chain,which amino acid may optionally be N-(1-6C)alkyl substituted; or J and Dtogether are the residue (R′)⁴(R′)⁵N—CH(R′)⁶—C(O)—.

[0068] Most preferred are the compounds of formula (XX) wherein J is H,(R′)²—SO₂—, (R′)⁷OOC—(CH(R′)³)_(p)—, (R′)¹ ₂N—CO—(CH(R′)³)_(p)—,

[0069] D is a bond, D-1-Tiq, D-3-Tiq, D-Atc, Aic, D-1-Piq, D-3-Piq or aD-amino acid having a hydrophobic side chain; or J and D together arethe residue (R′)⁴(R′)⁵N—CH(R′)⁶—C(O)—, wherein at least one of (R′)⁴ and(R′)⁵ is (R′)⁷OOC—(CH(R′)³)_(p)— or (R′)²—SO₂— and the otherindependently is (1-12C)alkyl, (2-12C)alkenyl, (2-12C)alkynyl,(3-12C)cycloalkyl, (7-15C)aralkyl, (R′)²—SO₂— or(R′)⁷OOC—(CH(R′)³)_(p)—, and (R′)⁶ is a hydrophobic side chain.

[0070] Preferably, the group Q in the compounds of formula (XX) has oneof the structures:

[0071] wherein X is O or S.

[0072] Like the compounds of formula (I), the compounds of formula (XX)have an anticoagulant effect and are useful for treating and preventingthrombin-mediated and thrombin-associated diseases, applicable as hereinbefore described.

[0073] In the description of the compounds of the formula (XX), thefollowing definitions are used. The term optionally substituted D,Lα-hydroxyacetyl means a group of the formula HO—CR^(a)R^(b)—C(O)—,wherein R^(a) and R^(b) independently are H, a hydrophobic side chain,or R^(a) and R^(b) together form a 5- or 6-membered ring, which isoptionally fused with one or two aliphatic or aromatic 6-membered rings,and which 5- or 6-membered ring consists of carbon atoms and optionallyone heteroatom selected from N, O and S. Preferred D,L α-hydroxyacetylgroups are 2-hydroxy-3-cyclohexyl-propionyl- and9-hydroxy-fluorene-9-carboxyl.

[0074] The term (1-12C)alkyl means a branched or unbranched alkyl grouphaving 1 to 12 carbon atoms, such as methyl, ethyl, t-butyl, isopentyl,heptyl, dodecyl, and the like. Preferred alkyl groups are (1-6C)alkylgroups, having 1-6 carbon atoms.

[0075] A (2-12C)alkenyl group is a branched or unbranched unsaturatedhydrocarbon group having 2 to 12 carbon atoms. Preferred are(2-6C)alkenyl groups. Examples are ethenyl, propenyl, allyl, and thelike.

[0076] The term (1-6C)alkylene means a branched or unbranched alkylenegroup having 1 to 6 carbon atoms, such as —(CH₂)_(b)— and b is 1 to 6,—CH(CH₃)—, —CH(CH₃)—(CH₂)—, etc.

[0077] A (2-12C)alkynyl group is a branched or unbranched hydrocarbongroup containing a triple bond and having 2 to 12 carbon atoms Preferredare (2-6C)alkynyl groups, such as ethynyl and propynyl.

[0078] A (6-14C)aryl group is an aromatic moiety of 6 to 14 carbonatoms. The aryl group may further contain one or more hetero atoms, suchas N, S, or O, also referred to as heteroaryl groups.

[0079] Examples of aryl groups are phenyl, naphthyl, (iso)quinolyl,indanyl, and the like. Most preferred is the phenyl group.

[0080] (7-15C)Aralkyl and (8-16C)aralkenyl groups are alkyl and alkenylgroups respectively, substituted by one or more aryl groups, the totalnumber of carbon atoms being 7 to 15 and 8 to 16, respectively.

[0081] The term (1-6C)alkoxy means an alkoxy group having 1-6 carbonatoms, the alkyl moiety of which having the meaning as previouslydefined.

[0082] The term (3-12C)cycloalkyl means a mono- or bicycloalkyl grouphaving 3-12 carbon atoms, being cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl, cyclo-octyl, the camphor group, etc..Cyclopentyl and cyclohexyl are preferred cycloalkyl groups.

[0083] The term halogen means fluorine, chlorine, bromine or iodine.

[0084] The term ester derivative means any appropriate ester derivative,preferably (1-4C)alkyl-esters, such as methyl-, ethyl- ort-butyl-esters.

[0085] The terms 1- and 3-Tiq mean 1,2,3,4-tetrahydroisoquinoline-1- and-3-carboxylic acid, respectively; 1- and 3-Piq areperhydroisoquinoline-1- and -3-carboxylic acid, respectively; Atc is2-aminotetralin-2-carboxylic acid; Aic is amino indane carboxylic acid;Phe is phenylalanine; Cha is cyclohexylalanine; Dpa is diphenylalanine;Coa is cyclooctylalanine; Chg is cyclohexylglycine; Nle is norleucine,Asp is aspartic acid.

[0086] The term hydrophobic side chain means a (1-12C)alkyl, optionallysubstituted with one or more (3-12C)cycloalkyl groups or (6-14C)arylgroups (which may contain a heteroatom, e.g. nitrogen) such ascyclohexyl, cyclo-octyl, phenyl, pyridinyl, naphthyl,tetrahydronaphthyl, and the like, which hydrophobic side chain mayoptionally be substituted with substituents such as halogen,trifluoromethyl, lower alkyl (for instance methyl or ethyl), loweralkoxy (for instance methoxy), phenyloxy, benzyloxy, and the like.

[0087] In the definitions, the term substituted in general means:substituted by one or more substituent. Amino acids having a basic sidechain are for example, but not limited to, arginine and lysine,preferably arginine. The term amino acids having a neutral side chainrefers to amino acids such as methionine sulphon and the like.

[0088] Cyclic amino acids are for example 2-azetidine carboxylic acid,proline, pipecolic acid, 1-amino-1-carboxy-(3-8C)cycloalkane (preferably4C, 5C or 6C), 4-piperidine carboxylic acid, 4-thiazolidine carboxylicacid, 3,4-dehydro-proline, azaproline, 2-octahydroindole carboxylicacid, and the like Preferred are 2-azetidine carboxylic acid, proline,pipecolic acid, 4-thiazolidine carboxylic acid, 3,4-dehydro-proline and2-octahydroindole carboxylic acid.

[0089] Also preferred serine protease inhibitors of the presentinvention are compounds of the formula (XXX):

[0090] in which formula Q is as previously defined; r is an integer of 0to 4; (R″)¹ is a hydrogen atom, a carboxyl group, an alkoxycarbonylgroup, a carboxyalkyl group, an alkoxycarbonylalkyl group, acarboxyalkoxy group or an alkoxycarbonylalkoxy group; (R″)² is ahydrogen atom, a hydroxyl group, a lower alkyl group or a lower alkoxygroup, M is an alkylene group having a carbon number of 1 to 4, whichmay have 1 or 2 substituents selected from the group consisting ofhydroxyalkyl, carboxyl, alkoxycarbonyl, carboxyalkyl andalkoxycarbonylalkyl; T is a single bond, an oxygen atom, a sulfur atomor a carbonyl group; K is a saturated or unsaturated 5- or 6-memberedheterocyclic moiety or cyclic hydrocarbon moiety optionally having asubstituent group, an amino group optionally having a substituent groupor an aminoalkyl group optionally having a substituent group.

[0091] Related compounds are known for instance from EP 0,540,051.According to the present invention the compounds of EP 0,540,051 arealtered by replacing the aromatic group carrying the amidine substituentby the group Q, thus improving in particular the absorptive propertiesof the compounds, such as examples having the structure:

[0092] Like the compounds of the structures (I) and (XX), the compoundsof structure (XXX) show a strong anticoagulant effect and are applicableas herein described.

[0093] In the compounds of the present invention represented by generalformula (XXX), any straight chain, branched chain or cyclic alkyl grouphaving 1 to 6 carbon atoms may be used as the lower alkyl group.Illustrative examples include methyl, ethyl, propyl, isopropyl, butyl,sec- or tert-butyl, pentyl, hexyl, cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl and the like. The lower alkoxy group may have 1 to 6 carbonatoms. Illustrative examples include methoxy, ethoxy, propoxy,isopropoxy, butoxy, sec- or tert-butoxy and the like. Thealkoxycarbonyl, carboxyalkyl, alkoxycarbonylkalkyl, carboxyalkoxy,alkoxycarbonylalkoxy and hydroxyalkyl groups preferably have 1 to 6carbon atoms, more preferably 1 to 4 carbon atoms, respectively.Illustrative examples of the alkoxycarbonyl group includemethoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, butoxycarbonyl and thelike. Illustrative examples of the carboxyalkyl group includecarboxymethyl, carboxyethyl, carboxypropyl and the like. Illustrativeexamples of the alkoxycarbonylalkyl group include methoxycarbonylmethyl,ethoxycarbonylmethyl, propoxycarbonylmethyl, methoxycarbonylethyl,ethoxycarbonylethyl, methoxycarbonylpropyl, ethoxycarbonylpropyl and thelike. Illustrative examples of the carboxylalkoxy group includecarboxymethoxy, carboxyethoxy, carboxypropoxy and the like. Illustrativeexamples of the alkoxycarbonylalkoxy group includemethoxycarbonylmethoxy, ethoxycarbonylmethoxy, propoxycarbonylmethoxy,methoxycarbonylethoxy, ethoxycarbonylethoxy and the like. Illustrativeexamples of the hydroxyalkyl group include hydroxymethyl, hydroxyethyl,hydroxypropyl, hydroxybutyl and the like. The saturated or unsaturated5- or 6-membered heterocyclic moiety may contain preferably one or twohetero-atom(s) selected from nitrogen and oxygen atoms. Illustrativeexamples of such a preferred type of heterocyclic rings includepyrrolidine, piperidine, imidazoline, piperazine, tetrahydrofuran,hexahydropyrimidine, pyrrole, imidazole, pyrazine, pyrrolidinone,piperidinone, morpholine and the like. More preferable are pyrrolidineand piperidine which contain one nitrogen atom as the hetero-atom.Illustrative examples of the saturated or unsaturated cyclic hydrocarbonmoiety include cyclopentyl, cyclohexyl and the like. Illustrativeexamples of the aminoalkyl group include aminomethyl, aminoethyl,aminopropyl and the like. Illustrative examples of the substituentsapplicable to these heterocyclic moieties and cyclic hydrocarbonmoieties include preferably lower alkyl, lower alkanoyl, carbamoyl,mono- or dialkylcarbamoyl, formimidoyl, alkanoimidoyl, benzimidoyl,carboxyl, alkoxycarbonylimino and the like, more preferably formimidoyland alkanoimidoyl groups. Illustrative examples of the substituentsapplicable to these amino groups and amino moieties of aminoalkyl groupsinclude preferably lower alkyl, pyradinyl, pyrrolidinyl, carbamoyl,mono- or dialkylcarbamoyl, lower alkanoyl, formimidoyl, alkanoimidoyl,benzimidoyl, alkoxycarbonyl and the like, more preferably pyrazinyi,pyrrolidinyl, formimidoyl, alkanoimidoyl groups. In this instance, eachof the alkyl, alkoxy, alkanoyl and the like listed above may preferablyhave a carbon number of from 1 to 6.

[0094] Other preferred serine protease inhibitors of the presentinvention are compounds of the formula (XL):

[0095] in which formula (R′″)¹ and (R′″)² are independently H, loweralkyl, aryl, heteroaryl, cycloalkyl or lower alkyl substituted by one ormore substituents selected from CONH₂, COO-(lower alkyl), aryl,heteroaryl and cycloalkyl; or (R′″)² is lower alkanoyl; (R′″)³ is H,COOH, CONH₂, COO-(lower alkyl), CONH-(lower alkyl) or CON(lower alkyl)₂;(R′″)⁴, (R′″)⁵ and (R′″)⁶ are independently H, lower alkyl, aryl,aralkyl or cycloalkyl; or (R′″)⁴ and/or one of (R′″)⁵ and (R′″)⁶ isheteroaryl or lower alkyl substituted with OH, SO₂H, SO₃H, guanidino,aryl-(lower alkoxy), lower alkoxy or lower alkylthio; or (R′″)² togetherwith (R′″)⁴ forms a tri- or tetramethylene group, in which (a) amethylene group may be replaced by S, SO or SO₂ or may be substitutedwith OH, lower alkyl, lower alkenyl or carboxy-(lower alkyl) or (b) oneof the methylene groups may be substituted with lower alkenyl and theother with (lower alkyl)-COOH; and at least one of (R′″)¹, (R′″)²,(R′″)⁴, (R′″)⁵ and (R′″)⁶ is the group of formula Q, Q having thepreviously defined meaning.

[0096] Related compounds are known from EP 0,728,758. The presentinvention is an improvement in the art when compared to these knowncompounds because of the presence of the specific group Q forintroducing the favourable properties as herein before described.

[0097] Like the compounds of the structures (I), (XX) and (XXX), thecompounds of structure (XL) show anticoagulant activity and areapplicable as herein described. The compounds of structure (XL) displaythis effect in particular through their specific thrombin- andFXa-inhibiting activity.

[0098] In the description of compounds of formula (XL) the term “lower”means a branched or unbranched group having 1-6 C-atoms. Preferred loweralkyl or lower alkanoyl groups contain up to 4 C-atoms, e.g. methyl,isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, and acetyl,respectively. “Aryl” alone or in combination means groups like phenyl,which may be substituted, for instance with amidino, guanidino,hydroxyamidino, nitro, amino or methylenedioxy. “Aralkyl” means an arylbound to a lower alkyl, e.g. a benzyl group, substituted in the phenylring, or phenylethyl. “Cycloalkyl” means saturated groups having 3-7C-atoms, like cyclohexyl. “Heteroaryl” means 5- to 10 membered aromaticgroups, which may consist of two rings, and contain one (or more)N-atom(s) and may be substituted, e.g. ,by one or more NH₂-groups. Anexample is chinazolinyl, such as 2,4-diaminochinazolin-6- or 7-yl.Examples of groups having amino, guanidino or N-hydroxyamidinosubstituents are amino-substituted chinazolinyl and (amino, amidino,guanidino or N-hydroxyamidino)-substituted phenyl, benzyl and loweralkyl groups.

[0099] Further preferred compounds according to the invention are theGpIIb/IIIa antagonists of the formula (L):

Q−[spacer]−COOH   (L),

[0100] wherein Q has the previously defined meaning; the distancebetween the amino substituted carbon atom of the group Q and the carbonatom of the carboxylate moiety has a length of 12-18 Å; the spacer isany suitable chemical moiety; and the carboxylate group may beesterified. The compounds of structure (L) show anticoagulant effect andare applicable as herein described.

[0101] From literature it is known, that a large group of spacers aresuitable for the above purpose, provided that the length thereof isrestricted (see Bioorg. & Med. Chemistry Letters, 7(2), 209-212, (1997),and references cited therein). Examples of compounds of formula (L) arederived from the following known compounds, in which the benzamidinemoiety has been replaced by the group Q (but not limited to theseexamples): the compounds Ro 43-8857 (J. Med. Chem. 35, 4393 (1992)), Ro44-9883, Ro 48-3657, Fradafiban, BIBL 12, FK-633, GR 144053, EMD 76 334,SR 121566, SB 208651, SC 54684, SC 54701, SC 52012, DMP 754, GPI 562 andcompounds described in EP 529,858, WO 96/20172, EP 496,378, EP 530,505,Bioorg. & Med Chem. 3, 539 (1995), WO 93/08174 and J.Am.Chem.Soc. 115,8861 (1993). This replacement by the group Q in the above compoundsresults in an improvement of the pharmacological properties of thecompounds, in particular of the absorptive properties in the intestines,as herein described.

[0102] Preferred compounds of formula (L) have the formula (La):

Q—C(O)—N(R″″)¹—U—C(R″″)¹(R″″)²—V   (La),

[0103] wherein Q has the previously defined meaning; (R″″)¹ isindependently H or (1-4C)alkyl; U is a bond, CH(R″″)¹; (R″″)² is(1-12C)alkyl, (2-12C)alkenyl, (2-12C)alkynyl, (3-8C)cycloalkyl,(6-14C)aryl, (7-15C)aralkyl or (8-16)aralkenyl, which may optionally besubstituted with (1-6C)alkyl, (3-8C)cycloalkyl, (1-6C)alkoxy, OH, COOH,CF₃ or halogen;V is a 5-, 6-, or 7-membered saturated, unsaturated oraromatic ring which may optionally contain one or more heteroatomsselected from O, N or S and which ring is substituted with one or twosubstituents selected from —(CH₂)_(v)—COO(R″″)¹ and—O—(CH₂)_(v)—COO(R″″)¹, v being 1, 2, 3 or 4.

[0104] More preferred compounds of formula (La) are those wherein m is0; U is a bond; (R″″)² is (1-4C)alkyl, phenyl or benzyl, which mayoptionally be substituted with OH or halogen and V is phenyl,piperidinyl, piperazinyl or thiazolyl, substituted with one substituentselected from —CH₂—COO(R″″)¹ and —O—CH₂—COO(R″″)¹.

[0105] Other preferred compounds of the formula (L) have the formula(Lb)

Q—Z¹—C(O)—N(R″″)³—Z²—[C(O)]_(y)—CH(R″″)⁴—(CH₂)_(z)—COO(R″″)   (Lb)

[0106] wherein Q has the previously defined meaning; Z¹ is a bond, C═Cor C≡C; (R″″)³ is H or (1-4C)alkyl; Z² is selected from

[0107] (R″″)⁴ is H, (1-12C)alkyl, (2-12C)alkenyl, (2-12C)alkynyl,(3-8C)cycloalkyl, (6-14C)aryl (7-15C)aralkyl or (8-16)aralkenyl, whichmay optionally be substituted with (1-6C)alkyl, (3-8C)cycloalkyl,(1-6C)alkoxy, OH, COOH, CF₃ or halogen; y is 0 or 1 and z is 0 or 1.

[0108] More preferred are the compounds of formula (Lb) wherein Z¹ isC═C; Z² is

[0109] (R″″)⁴ is H; y is 1 and z is 0.

[0110] Other preferred compounds of the formula (L) have the formula(Lc)

Q—N(H)—C(O)—F—C(O)—N(R″″)⁵—(CH₂)_(f)—N(R″″)⁵—CH₂COOH   (Lc),

[0111] wherein Q has the previously defined meaning;

[0112] (R″″)⁵ is independently H, (1-4C)alkyl or benzy or both (R″″)⁵groups are an ethylene bridge to form a 6- or 7-membered ring togetherwith N—(CH₂)_(u)—N to which they are bound;

[0113] F is C═C, or 1,2-, 1,3- or 1,4-phenylene, or1,2-(4-5C)heteroarylene, 2,3-naphthylene or 1,2-(5-7C)cycloalkylene,which groups may optionally be substituted with (1-4C)alkyl;

[0114] and f is 2 or 3.

[0115] The terms used in the definitions of the compounds of formula(L), (La), (Lb) and (Lc) have the same meaning as those used for thecompounds of formula (XX).

[0116] The present invention further relates to the finding that also inother therapeutic compounds the presence of the group Q gives rise to animproved pharmacological profile. Especially when Q is used to replace abasic moiety in compounds of which is known that they require such amoiety for their therapeutic activity, an improvement of the absorptiveproperties is observed, in particular when that basic moiety is a(hetero)arylguanidino or (hetero)arylamidino moiety. Preferredtherapeutic compounds of the invention have an apparent Caco-2permeability of 8 nm/s or higher. Preferably the group Q has the formula

[0117] m being as previously defined.

[0118] The compounds of the invention may be used in a broad range oftherapeutic applications which require oral administration of a drug orwherein oral administration thereof is considered favourable, such as inparticular (but not limited to) CNS-active compounds, compounds usefulfor treating immunological disorders, antithrombotic agents, and thelike. Preferred compounds of the present invention are antithromboticagents.

[0119] The term “therapeutic compound” as used herein means any compoundwhich can be used in therapy, which implies the curing of a disease ormalfunction of the body and which covers prophylactic treatment.

[0120] Compounds of the present invention wherein Q has the formula

[0121] wherein the substructure

[0122] is a structure selected from

[0123] wherein X is O or S;

[0124] X′ being independently CH or N; and m is 0, 1, 2 or 3; can beprepared using suitable starting compounds and methods as described inthe literature e.g. 4-amino-6-chloroquinazoline and4-amino-7-chloroquinazoline as described by A. Rosowsky and N.Papathanasopoulos in J. Heterocycl. Chem. 9, 1235 (1972);4-aminothieno[2,3d]pyridazine or 7-aminothieno[2,3d]pyridazine by M.Robba, B. Roques and Y. Le Guen in Bull. Soc. Chim. France 4220, (1967);4-aminothieno[2,3d]pyrimidine by M. Robba, J. -M. Lecompte and M. Cugnonde Sevricourt in Bull. Soc. Chim. France 592, (1975);4-aminothieno[3,2d]pyrimidine by M. Robba, J. -M. Lecompte and M. Cugnonde Sevricourt in Tetrahedron 27, 487, (1971);4-amino-6-bromothieno[2,3d]pyrimidine by M. Robba, J. -M. Lecompte andM. Cugnon de Sevricourt in Bull. Soc. Chim. France 761, (1976);4-amino-6-bromoquinazoline by M. F. G. Stevens and A. Kreutzberger inAngew. Chem. 81, 84, (1969).

[0125] The compounds of the present invention wherein Q has the formula

[0126] wherein m is 0, 1, 2 or 3 and X is CH═CH, O or S, can be preparedusing compounds of formula (II).

[0127] A suitable method starts from a compound of formula (III)(wherein the halogen atom, if present, preferably is Br), the hydroxygroup of which is chlorinated, e.g. by treatment with POCl₃, to give thecompound of formula (IV), followed by conversion into the amino analogueof formula (II) , for example by first converting the chloro-substituentinto a phenoxy-substituent by reaction with phenol under alkalineconditions, and subsequently treatment with ammonium acetate, or bydirect conversion of the chloro-compound into the correspondingamino-compound by heating the former with ammonia under pressure.

[0128] The compound of formula (IV) in this sequence, wherein X is CH═CHand the halogen is Br, may also be prepared by converting thecorresponding unsubstituted compound into the N-oxide salt, e.g. with aperacid, such as m-chloroperbenzoic acid, followed by HCl treatment, andsubsequently reacting this N-oxide salt with a chlorinating reagent,like POCl₃.

[0129] Compounds according to formula (I) can be prepared bydeprotection of the following compound (e.g. by saponification)

R¹—Y—[NR²—A—C(O)]_(n)—NR³—CHR⁴—C(O)—O—G,

[0130] wherein G e.g. is an alkyl or benzyl group, followed by couplingwith R⁵—H, or they can be prepared by deprotection of the compound

Pg—NR³—CHR⁴—C(O)R⁵,

[0131] wherein Pg is an N-protecting group, followed by coupling withone of the groups

R¹—Y—NR²—A—C(O)—OH, R¹—Y—Lg

[0132] or

R¹—C(O)—OH,

[0133] wherein Lg is a leaving group.

[0134] Compounds of formula (I) wherein R⁴ is Q may be prepared startingwith a compound of formula (V),

[0135] or a derivative thereof wherein the amino group at the aromaticgroup (arylamino) is protected, wherein X and R³ have the previouslydefined meanings. The carboxylic acid group of a compound of formula (V)is esterified, e.g. by treatment with R⁶OH and thionyl chloride, whereinR⁶ has the previously defined meaning, to form a compound of formula(VI), or an arylamino protected derivative thereof.

[0136] A compound of formula (VI) is converted into a compound offormula (Ia), being the compound of formula (I) wherein R⁵ is OR⁶, or anarylamino protected derivative thereof, either by coupling to R¹YLg (Lgis a leaving group) (n=0), for example by reaction with R¹YCl underbasic conditions, e.g. by using triethylamine, or by peptide couplingwith R¹YNR²AC(O)OH (n=1) or R¹C(O)OH (n=0) using as a coupling reagentfor example N,N-dicyclohexylcarbodiimide (DCCI) and1-hydroxybenzotriazole (HOBT) or2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate(TBTU), wherein R¹, R², Y, n and A have the previously defined meanings.The protective group, if present, may optionally be removed.

[0137] The ester group of a compound of formula (Ia) or an arylaminoprotected derivative thereof may be saponified to form the correspondingacid, after which the protective group, if present, may optionally beremoved. The acid formed from a compound of formula (Ia) may be coupledto an amine of formula HNR⁷R⁸, wherein R⁷ and R⁸ have the previouslydefined meanings, for example by using DCCI/HOBT or TBTU, followed byremoval of the optionally present protective group, to give a compoundof formula (Ib), which is the compound of formula (I), wherein R⁵ isNR⁷R⁸.

[0138] An alternative procedure for the preparation of a compound offormula (Ib) starts with the protection of the N-terminus of a compoundof formula (V) with an N-protecting group Pg, such as Boc(tert-butoxycarbonyl), forming a compound of formula (VII), where alsothe arylamino group may be protected.

[0139] Subsequently, a compound of formula (VII) is coupled to an amineof formula HNR⁷R⁸ according to the procedure as described above for theconversion of a compound of formula (Ia) into (Ib), after which theN-terminus is deprotected, followed by a coupling to R¹YLg (n=0), or bypeptide coupling with R¹YNR²AC(O)OH (n=1) or R¹C(O)OH (n=0) according tothe procedures as described above for the conversion of a compound offormula (VI) into a compound of formula (Ia), followed by removal of theoptionally present protective group, giving the compound of formula(Ib).

[0140] A suitable process for the preparation of the compound of formula(V) is the conversion of a compound of formula (VIII), wherein Pg is anN-protecting group and Lg is a leaving group, such as the mesyl group,into a compound of formula (IX), for example by reaction with anappropriate amino acid derivative of the general formulaPgR³N—CH[C(O)OR]₂ in the presence of a base, e.g[[(1,1-dimethylethoxy)carbonyl]amino] propanedioic acid diethyl ester inthe presence of sodium ethoxide, wherein Pg and R³ have the previouslydefined meanings and both Pg-groups may be the same or different, and Ris a branched or unbranched (1-8C)alkyl group, such as ethyl. (If hereR³ is H, the amino group may be alkylated later in the procedureaccording to methods well known in the art to form compounds wherein R³is alkyl.) Hydrolysis and decarboxylation of a compound of formula (IX)gives a compound of formula (V).

[0141] The compound according to formula (VIII) may be prepared bymethods known in the art. The arylamino group of a compound of formula(II) is protected with an N-protecting group, after which the ringwherein X is located is provided with a formyl substituent either bytreatment with a base, like lithium diisopropylamide, or with anorganometallic reagent, like n-butyllithium, followed by addition ofN,N-dimethylformamide, forming a compound of formula (X). (For compoundswherein m is 2 or 3, appropriate analogues of a compound of formula (X)may be prepared by this method which subsequently may be convertedaccording the procedures as described for the conversion of a compoundof formula (X).)

[0142] The compound of formula (X) is reduced, e.g. using sodiumborohydride, into the corresponding alcohol, which then is convertedinto a leaving group, e.g. an appropriate sulfonate group, therebyforming a compound of formula (VIII).

[0143] Compounds of formula (X) are suitable intermediates for thepreparation of compounds of formula (I) wherein R³ is Q. In thosepreparations a compound of formula (X) is reacted with an appropriateamino acid derivative of the general formula NH₂—CHR⁴—C(O)OR, R⁴ and Rhaving the previously defined meanings. The resulting imine is eitherdirectly converted into a compound of formula (XI) by reduction, usinge.g. sodium cyanoborohydride, or isolated and subsequently reduced withan suitable reducing agent like sodium borohydride to form a compound offormula (XI).

[0144] An alternative procedure for the preparation of a compound offormula (XI) starts from a compound of formula (VIII) by substitution ofLg by an appropriate amino acid derivative of the general formulaNH₂—CHR⁴—C(O)OR.

[0145] Compounds of formula (XI) are then converted into compounds offormula (Ic), the compounds of formula (I) wherein R³ is Q and R⁵ isOR⁶, by a procedure analogous to the preparation of compounds of formula(Ia) from compounds of formula (VI) followed by removal of theprotective group.

[0146] From compounds of formula (Ic) compounds of formula (Id) areprepared, which are the compounds of formula (I), wherein R³ is Q and R⁵is NR⁷R⁸, following the procedure as described for the conversion of(Ia) into (Ib).

[0147] Alternatively, a compound of formula (Id) may be obtained afterprotection of the N-terminus of (XI) with an N-protecting group Pg, suchas Boc, both protecting groups in the molecule being the same ordifferent, and saponification of the ester group to give intermediate(XII), and further derivatization as described for the conversion of acompound of formula (VII) into a compound of formula (Ib).

[0148] Compounds of formula (I) wherein R¹² together with R³ is—(CH₂)_(s)— can be prepared from amines of formula (VI) wherein R³ ishydrogen and the arylamino-is optionally protected. Reaction of theseamines with aldehydes of formula Pg(R²)NCH((CH₂)_((s-1))CHO)COOG,wherein Pg is a N-protecting group and R², s and G have the previouslydefined meanings, gives imides which are reduced using e.g. sodiumcyanoborohydride to give cyclic compounds of formula (XIII). Afterremoval of the N-protecting group Pg the moiety,R¹Y can be introduced asdescribed for the conversion of compounds of formula (VI) into compoundsof formula (Ia). The group OR⁶ can be modified in the same way asdescribed for the conversion of compounds of formula (Ia) into compoundsof formula (Ib) and removal of protecting groups if present givescompounds of formula (Ie), Furthermore, compounds of formula (Ie) and(XIII) can be prepared using the methods described by H. Mack et al. inJ. Enzyme Inhibition 9, 73 (1995), wherein instead of the cyanophenylbuilding blocks used in the literature, building blocks containingmoiety Q or a arylamino protected derivative thereof (e.g. compounds offormula (VI)) can be used.

[0149] Compounds of formula (XX) can be prepared from compounds offormula (X). Reaction of a compound of formula (X) with an amine offormula (R′)¹NH₂ gives an imide, which is either directly converted intoa compound of formula (XXI) by reduction, using e.g. sodium cyanoborohydride, or is isolated and subsequently reduced with a suitableagent like sodium borohydride to form a compound of formula (XXI),wherein (R′)¹ has the previously defined meanings.

[0150] Alternatively a compound of formula (X) is reduced, using e.g.sodium borohydride, into the coresponding alcohol which then isconverted into an azide using the method described by A. S. Thompson etal. in J. Org. Chem. 58, 5886 (1993). This azide can be reduced into anamine using reagents known in the art such as Pd/CaCO₃ catalyzedhydrogenation to yield a compound of formula (XXI) in which (R′)¹ ishydrogen.

[0151] A compound of formula (XXI), or a derivative thereof wherin thearylamino is not protected, can be coupled with carboxylate compounds offormula J—D—E—OH, in which J, D and E have the previously definedmeanings, or a protected derivative thereof, using peptide couplingmethods. Subsequent removal of the optionally present protective groupsgives compounds of formula (XX).

[0152] Compounds of formula (XXa) can be prepared by reaction of aminesof formula (XXI), wherein (R′)¹ is hydrogen with aldehydes of formulaPgHNCH((CH₂)_((u-1))CHO)COOG, wherein Pg is a N-protecting group and uand G have the previously defined meanings, using the method describedfor compounds of formula (Ie).

[0153] Compounds of formula (XXX) may be prepared in several ways inwhich the synthetic connection of moiety Q and the substituted phenylpart is made in moiety M. The method described in EP 0540051 may be usedto prepare componds of formula (XXX) starting with compounds of formula(VIII).

[0154] Compounds of formula (XL) can be prepared by reaction of acompound of formula (XLI) with a compound of formula (XLII) and acompound of formula (XLIII), wherein (R′″)¹, (R′″)², , (R′″)⁵, and(R′″)⁶ have the previously defined meanings according to the methodsdescribed in EP 0728758. When (R′″)¹ or (R′″)² is the group of formula Qcompounds of formula (II), (VIII) or (X) or an arylamino protectedderivative thereof can be used to prepare compounds of formula (XLI) or(XLII). When (R′″)⁴ is the group of formula Q amino acids of formula(VI) or an arylamino protected derivative thereof can be used asstarting materials, When (R′″)⁵ or (R′″)⁶ is the group of formula Q,compounds of formula (X) or an arylamino protected derivative thereofcan be used as a starting material, being aldehydes of formula (XLIII),or used to prepare ketones of formula (XLIII).

[0155] Compounds of formula (L) can be prepared using methods known inthe art described for Gp IIb/IIIa receptor antagonists containing a(hetero)arylamidine moiety instead of moiety Q. Compounds of formula(La) can conveniently be prepared from compounds of formula (X). Analdehyde of formula (X) is oxidized, using e.g. sodium chlorite, intothe corresponding carboxylic acid. This carboxylic acid, or a derivativethereof wherein at the arylamino is not protected, can be coupled withan amine linker of formula HN(R″″)¹—U—(R″″)¹(R″″)²—V or a N-protectedderivative thereof, wherein (R″″)¹, (R″″)²-and V have the previouslydefined meanings, using peptide coupling methods (e.g. using the aminesand methods described in J. Med. Chem. 35, 4393 (1992), EP 0,505,868 orJ. Med. Chem. 39, 3193 (1996)). Subsequent removal of the optionallypresent protective groups gives compounds of formula (La). Using asimilar peptide coupling conditions a linker molecule having one freecarboxylate can also be coupled with compound of formula (XXI), or aderivative thereof wherein the arylamino is not protected. Subsequentremoval of the optionally present protective groups gives compounds offormula (L). Also halogen containing compounds of formula (II) can beused as starting material. The linker molecule can be attached using Pdmediated reactions such as Suzuki coupling, Heck reaction, or firsttransmetalation, using e.g. n-BuLi, and secondly reaction with a linkermolecule containing an electrophilic function. Subsequent removal of theoptionally present protective groups gives compounds of formula (L). Forinstance, Heck reaction of a halogen containing compound of formula (II)and an acrylic acid derivative leads to compounds of formula (Lb). Analternative method for the preparation of compounds of formula (Lb) is acondensation reaction of aldehydes of formula (X) with a malonic acidderivative. Modification of this malonic acid derivative using methodsknown in the art and subsequent removal of optionally present protectinggroups yields compounds of formula (Lb).

[0156] Compounds of the invention can also be prepared using a solidphase synthesis strategy. To prepare componds of formula (Ia) thecarboxylic acid of the compounds of formula (VII) can be covalentlyattached to a polymeric support such as a polystyrene-resin using aester or amide bond as anchoring bond. In this case protection of thearylamino function is preferred. For example anchoring using a esterbond to the Kaiser oxime resin, Boc-protection of the N-terminus andacetyl amide protection of the arylamino group. The N-terminusprotecting group can be removed selectively followed by coupling toR¹YLg (n=0), or by peptide coupling with R¹C(O)OH (n=0), Pg²NR²AC(O)OH(n=1) or R¹YNR²AC(O)OH (n=1) according to the procedures described abovefor the conversion of compounds of formula (VI) into compounds offormula (Ia). Protecting group Pg² is a protecting group that can beremoved selectively. Removal of Pg² liberates the N-terminus which canbe coupled with R¹YLg (n=0), or with R¹C(O)OH (n=0) by peptide couplingmethods according to the procedures described above for the conversionof compounds of formula (VI) into compounds of formula (Ia). Cleavage ofthe anchoring bond and removal of protecting groups, if present, givescompounds of formula (Ia) or (Ib) depending on the type of anchoringbond and way of cleavage used. For example cleavage of the anchoringester bond to the Kaiser oxime resin with amines of type HNR⁷R⁸ followedby removal of the optionally present protective groups yields compoundsof formula (Ib).

[0157] The solid phase synthesis strategy outlined above starting fromcompounds of formula (VII) can also be applied using the carboxylic acidof compounds of formula (XII) to yield compounds of formula (Ic) or(Id). Further, this strategy can be applied to carboxylic acids offormula J—D—E—OH— in which J, D, and E have the previously definedmeanings—, a synthetic precursor thereof or a protected derivativethereof. Cleavage of the anchoring bond to the Kaiser oxime resin usingamines of formula (XXI) yields compounds of formula (XX) after removalof the optionally present protecting group.

[0158] Alternatively, the arylamino functionality of moiety Q can beused to be covalently attached to a polymeric support using e.g. acarbamate functionality as method of anchoring and used in a solid phasesynthesis.

[0159] Several solid-phase synthesis strategies especially in thesolid-phase synthesis of peptides are known in the art. An overview ofsolid-phase peptide synthesis is given by P. Lloyd-Williams, F.Albericio and E. Giralt in Tetrahedron 48, 11065-11133 (1993).

[0160] The peptide coupling, as mentioned as a procedural step in theabove described method to prepare the compounds of the invention, can becarried out by methods commonly known in the art for the coupling—orcondensation—of peptide fragments such as by the azide method, mixedanhydride method, activated ester method, or, preferably, by thecarbodiimide method, especially with the addition of catalytic andracemisation suppressing compounds like N-hydroxy-succinimide andN-hydroxybenzotriazole. An overview is given in The Peptides, Analysis.Synthesis, Biology, Vol 3, E. Gross and J. Meienhofer, eds. (AcademicPress, New York, 1981).

[0161] The term N-protecting group as used in this whole document meansa group commonly used in peptide chemistry for the protection of anα-amino group, like the tert-butyloxycarbonyl (Boc) group, thebenzyloxycarbonyl (Z) group, the 9-fluorenylmethyloxycarbonyl (Fmoc)group or the phthaloyl (Phth) group. Removal of the protecting groupscan take place in different ways, depending on the nature of thoseprotecting groups. Usually deprotection takes place under acidicconditions and in the presence of scavengers. An overview of aminoprotecting groups and methods for their removal is given in the abovementioned The Peptides, Analysis, Synthesis Biology, Vol 3.

[0162] Suitable leaving groups (Lg) are known in the art, for examplefrom A. L. Ternay: Contemporary Organic Chemistry (2^(nd) ed., W. B.Saunders Company, 1979, see pages 158 and 170-172). Preferred leavinggroups are chloride, mesylate and tosylate.

[0163] The compounds of the invention, which can occur in the form of afree base, may be isolated from the reaction mixture in the form of apharmaceutically acceptable salt. The pharmaceutically acceptable saltsmay also be obtained by treating the free base of formula I with anorganic or inorganic acid such as HCl, HBr, HI, H₂SO₄, H₃PO₄, aceticacid, propionic acid, glycolic acid, maleic acid, malonic acid,methanesulphonic acid, fumaric acid, succinic acid, tartaric acid,citric acid, benzoic acid, ascorbic acid and the like.

[0164] The compounds of this invention may possess one or more chiralcarbon atoms, and may therefore be obtained as a pure enantiomer, or asa mixture of enantiomers, or as a mixture containing diastereomers.Methods for obtaining the pure enantiomers are well known in the art,e.g. crystallization of salts which are obtained from optically activeacids and the racemic mixture, or chromatography using chiral columns.

[0165] The compounds of the invention may be administered enterally orparenterally. The exact dose and regimen of these compounds andcompositions thereof will neccessarily be dependent upon the needs ofthe individual subject to whom the medicament is being administered, thedegree of affliction or need and the judgment of the medicalpractitioner. In general parenteral administration requires lowerdosages than other methods of administration which are more dependentupon absorption. However, the daily dosages are for humans preferably0.001-100 mg per kg body weight, more preferably 0.01-10 mg per kg bodyweight.

[0166] The medicament manufactured with the compounds of this inventionmay also be used as adjuvant in acute anticoagulant therapy. In such acase, the medicament is administered with other compounds useful intreating such disease states.

[0167] The compounds may also be used with implantable pharmaceuticaldevices such as those described in U.S. Pat. No. 4,767,628, the contentsof which are incorporated by this reference. Then the device willcontain sufficient amounts of compound to slowly release the compound(e.g. for more than a month).

[0168] Mixed with pharmaceutically suitable auxiliaries, e.g. asdescribed in the standard reference, Gennaro et al., Remington'sPharmaceutical Sciences, (18th ed., Mack Publishing Company, 1990, seeespecially Part 8: Pharmaceutical Preparations and Their Manufacture)the compounds may be compressed into solid dosage units, such as pills,tablets, or be processed into capsules or suppositories. By means ofpharmaceutically suitable liquids the compounds can also be applied inthe form of a solution, suspension, emulsion, e.g. for use as aninjection preparation, or as a spray, e.g. for use as a nasal spray.

[0169] For making dosage units, e.g. tablets, the use of conventionaladditives such as fillers, colorants, polymeric binders and the like iscontemplated. In general any pharmaceutically acceptable additive whichdoes not interfere with the function of the active compounds can beused.

[0170] Suitable carriers with which the compositions can be administeredinclude lactose, starch, cellulose derivatives and the like, or mixturesthereof, used in suitable amounts.

[0171] The invention is further illustrated by the following examples.

EXAMPLES

[0172] Melting points were measured on a Buchi 535 and are uncorrected.The ¹H N measurements were performed on BRUKER AC 200, BRUKER AM 360 andBRUKER DRX 400 spectrophotometers operating at a ¹H frequency of 200,360 and 400 MHz respectively. The ¹⁹F measurements were performed on a‘BRUKER AC 200 spectrophotometer operating at a ¹⁹F frequency of 188MHz.

[0173] 6-Aiq: [[(1-amino-6-isoquinolinyl)methyl]amino]

[0174] Atp: [[(4-aminothieno[3,2c]pyridin-2-yl)methyl]amino]

[0175] Azt: azetidine-2-carboxylate

[0176] Boc: t-butyloxycarbonyl

[0177] Cha: cyclohexylalanyl

[0178] Gly: glycyl

[0179] Phe: phenylalanyl

[0180] Pro: prolyl

[0181] TFA: trifluoroacetic acid

Example 13-(1-amino-6-isoquinolinyl)-2-[[2-[(2-naphthalenylsulfonyl)amino]acetyl]amino]propionicacid methyl ester hydrochloride (1k)

[0182] 1a. 6-Bromoisoquinoline N-oxide hydrochloride

[0183] To a stirred solution of 13.2 g of 6-bromoisoquinoline (Tyson, F.L., J. Am. Chem. Soc. 61, 183 (1939)) in 250 mL of dichloromethane atroom temperature was added in portions 16.2 g of m-chloroperbenzoic acid(purity 70%). After stirring the mixture for one hour 200 mL of methanolwas added and the bulk was reduced to 150 mL. A hydrogen chloridesolution in methanol (100 mL, 0.75 M) was added and after slightlyheating a clear solution was obtained. Diethylether (250 mL) was addedto this solution and cooled in an ice bath. The hydrochloride saltprecipitated and was isolated by filtration to give 15 g of 1a. M.p.194-196° C.

[0184] 1b. 6-Bromo-1-chloroisoquinoline

[0185] 70 mL of phosphorylchloride were added to 14.9 g of 1a and themixture was heated at 90° C. for 2 hours. Excess of phosphorylchloridewas evaporated, water was added, followed by aqueous 2N NaOH until pH 9and the mixture was extracted with dichloromethane. The dichloromethaneextract was washed with brine, dried (MgSO₄) and concentrated. Theresidue was chromatographed on silica gel (toluene/ethyl acetate=4/1)giving 10.1 g of 1b. M.p.=102.7-103.7° C.

[0186] 1c. 6-Bromo-1-phenoxyisoquinoline

[0187] A mixture of 10 g of 1b, 31 g of phenol and 4 g of KOH was heatedat 140° C. for 2 hours. After cooling to room temperature aqueous 3NNaOH was added and the mixture was extracted with dichloromethane. Thedichloromethane extract was washed with aqueous 2N NaOH, washed withwater, dried (MgSO₄) and concentrated to yield 12.2 g of 1c. ¹H—NMR 200MHz (CDCl₃) δ: 7.1-7.6 (6H, m), 7.70 (1H, dd, J=9 Hz and J=2 Hz),7.95-8.10 (2H, m), 8.31 (1H, d, J=9 Hz).

[0188] 1d. 1-Amino-6-bromoisoquinoline

[0189] A mixture of 12 g of 1c and 27 g of ammonium acetate was heatedat 150° C. for 14 hours. After cooling to room temperature aqueous 3NNaOH was added and the mixture was extracted with ethyl acetate. Theethyl acetate extract was washed with brine and aqueous 2N hydrochloricacid was added until pH2-3. The acid aqueous layer was separated, madebasic (pH10) with aqueous 2N NaOH and extracted with ethyl acetate. Theethyl acetate layer was washed with brine, dried (MgSO₄) andconcentrated to give 6.4 g of 1d. ¹H—NMR 200 MHz (CDCl₃) δ: 5.1 (2H, brs), 6.96 (1H, dd, J=6 Hz and J=1 Hz), 7.54-7.70 (2H, m), 7.88 (1H, d,J=2 Hz), 7.98 (1H, d J=6 Hz).

[0190] 1e. N-(6-bromo-1-isoquinolinyl)benzamide

[0191] 7.0 g of benzoic anhydride was added to a solution of 6.38 g of1d in 70 mL of pyridine at room temperature and the solution was heatedat 125° C. for 1 hour. The pyridine was evaporated and water anddichloromethane were added. The organic layer was separated, washed withwater, dried (MgSO₄) and concentrated. The crude product was dissolvedin dichloromethane and diethyl ether was added to give an amorphoussolid that was isolated by filtration (yield 7.84 g). The filtrate waschromatographed on a silica gel column (dichloromethane/methanol: 95/5)giving an additional 1.0 g of 1e. ¹H—NMR 200 MHz (CDCl₃) δ: 6.90 (1H,d,J=6 Hz), 7.35-8.48 (9H, m), 8.87 (1H, d, J=9 Hz).

[0192] 1f. N-(6-formyl-1-isoquinolinyl)benzamide

[0193] To a stirred solution of 29 mL of n-BuLi (1.6 M in hexane) in 40mL of dry tetrahydrofuran under a nitrogen atmosphere at −78° C. wasadded dropwise a solution of 2.5 g of 1e in 60 mL of dry tetrahydrofuranover a period of 15 min. After stirring for 30 min a mixture of 34 mL ofN,N-dimethylformamide and 20 mL of tetrahydrofuran was added fast. Thecooling bath was removed, the reaction mixture was allowed to come to 0°C. and was poured into ice cold aqueous hydrochloric acid (60 mL, 0.5N). The mixture was adjusted to pH 6, brine was added and the mixturewas extracted with ethyl acetate. The ethyl acetate extract was dried(MgSO₄) and concentrated under reduced pressure. Purification on silicagel (toluene/ethyl acetate: 3/1) gave 1.3 g of aldehyde If. ¹H—NMR 200MHz (CDCl₃) δ: 7.12 (1H, d, J=7 Hz). 7.40-8.51 (9H, m), 9.26 (1H, d, J=8Hz), 10.23 (1H, s).

[0194] 1g. N-[6-(hydroxymethyl)-1-isoquinolinyl]benzamide

[0195] To a stirred suspension of 1.23 g of 1f in 40 mL oftetrahydrofuran and 15 mL of methanol was added 217 mg sodiumborohydride in small portions. After stirring the mixture at ambienttemperature for 5 min, 50 mL of water was added and tetrahydrofuran andmethanol were removed in vacuo. Brine was added and the mixture wasextracted with ethyl acetate. The ethyl acetate extract was dried(MgSO₄) and concentrated under reduced pressure giving 1.27 g of 1g.¹H—NMR 200 MHz (CDCl₃) δ: 4.75 (2H, s), 6.95 (1H, br.s), 7.30-7.55 (7H,m), 8.25-8.38 (2H, m), 8.69 (1H, br.s).

[0196] 1h.[[1-(benzoylamino)-6-isoquinolinyl]methyl][[(1,1-dimethylethoxy)carbonyl]amino]propane-dioicacid diethyl ester

[0197] To a stirred suspension of 1.27 g of 1g in 30 mL ofdichloromethane at 0° C. was added 1.23 mL of triethylamine and 0.69 mLof methane sulfonylchloride and the mixture was allowed to warm to roomtemperature. After stirring for 2 hours, 40 mL of tetrahydrofuran and1.22 g of lithium chloride were added and the suspension was stirred for16 hours at room temperature. Brine was added and the organic layer wasseparated. The aqueous layer was extracted with dichloromethane. Thecombined organic layers were dried over magnesium sulfate andconcentrated under reduced pressure keeping the temperature below 30° C.The residue was coevaporated with toluene under reduced pressure againkeeping the temperature below 30° C. The residue (chloride) wasimmediately dissolved in 20 mL of dioxane and added to sodium malonatereaction mixture A. [This sodium malonate reaction mixture A wasobtained by addition of 3 g of[[(1,1-dimethylethoxy)carbonyl]amino]propanedioic acid diethyl ester(Paik, Y. H., Dowd, P., J. Org. Chem. 51, 2910-2913 (1986)) in 10 mL ofdioxane to a solution of sodium ethoxide (10 mmol) in 10 mL of dioxaneand 30 mL of ethanol, stirring for 10 min at room temperature andsubsequently addition of 0.5 g of sodium iodide.] After addition of thecrude chloride the reaction mixture was stirred at 80° C. for two hours.After cooling to room temperature water was added, the mixture wasneutralized with aqueous 1N hydrochloric acid and extracted with ethylacetate. The organic layer was dried (MgSO₄) and concentrated underreduced pressure. The residue was chromatographed on silica gel(toluene/ethyl acetate: 5/1) giving 1.45 g of 1h. 1H—NMR 200 MHz (CDCl₃)δ: 1.31 (6H, t), 1.51 (9H, s), 3.83 (2H, s), 4.18-4.39 (4H, m), 5.77(1H, s), 6.92 (1H, d, J=7 Hz), 7.30-7.56 (6H, m), 8.41-8.49 (2H, m),8.92 (1H, d, J=9 Hz).

[0198] 1i. 2-Amino-3-(1-amino-6-isoquinolinyl)propionic aciddihydrochloride

[0199] 15 mL of acetic acid and 30 mL of a solution of 6N hydrochloricacid were added to 1.34 g of 1h and heated at 100° C. for 16 h. Aftercooling to room temperature the aqueous solution was extracted twicewith diethyl ether. The aqueous phase was concentrated under reducedpressure giving 0.79 g of amino acid 1i. ¹H—NMR 200 MHz (D₂O) δ:3.33-3.56 (2H, m), 4.36 (1H, dd, J=6.5 Hz and J=7.5 Hz), 7.12 (1H, d,J=7 Hz), 7.46 (1H, d, J=7 Hz), 7.62 (1H, dd, J=8 Hz and J=2 Hz), 7.73(1H, d, J=2 Hz), 8.18 (1H, d, J=8 Hz).

[0200] 1j. 2-Amino-3-(1-amino-6-isoquinolinyl)propionic acid methylester dihydrochloride

[0201] 1.1 mL of thionyl chloride was added to a cooled solution (icebath) of 65 mg of amino acid 1i in 4 mL of methanol. After stirring at50° C. for 3 hours the mixture was concentrated under reduced pressureto give 66 mg of methyl ester 1j. The crude product was used withoutfurther purification. ¹H—NMR 200 MHz (CD₃OD) δ: 3.36-3.59 (2H, m), 3.81(3H, s), 4.53 (1H, t, J=7 Hz), 7.23 (1H, d, J=7 Hz), 7.60 (1H, d, J=7Hz), 7.73 (1H, dd, J=9 Hz and J=2 Hz) 7.87 (1H, d, J=2 Hz), 8.47 (1H, d,J=9 Hz),

[0202] 1k.3-(1-amino-6-isoquinolinyl)-2-[[2-[(2-naphthalenylsulfonyl)amino]acetyl]amino]propionicacid methyl ester hydrochloride

[0203] 62 mg of 1j was coevaporated with N,N-dimethylformamide and 3 mLof N,N-dimethyl-formamide, 58 mg of2-[(2-naphthalenylsulfonyl)amino]acetic acid (WO 92/16549), and 0.1 mLof N-ethylmorpholine (pH of the mixture was 8) were added. The mixturewas cooled at 0° C. and 47 mg of 1-hydroxybenzotriazole (HOBt) and 48 mgof N,N-dicyclohexylcarbodiimide (DCC) were added. After stirring for 16hours at room temperature the mixture was concentrated in vacuo. Waterwas added, the pH was adjusted to 8-9 and extracted with ethyl acetate.The ethyl acetate extract was dried (MgSO₄), concentrated andchromatographed on silica gel (ethyl acetate/pyridine/acetic acid/water:81/31/18/7). The product was dissolved in ethyl acetate, washed withwater (adjusted to pH 8-9), dried (MgSO₄) and concentrated to give 43 mgof free base. One equivalent hydrochloric acid was added andlyophilisation (t-butanol/water) afforded 1k. ¹H—NMR 200 MHz (CD₃OD) δ:3.09-3.41 (2H, m), 3.55 (2H, s), 3.69 (3H, s), 4.77 (1H, dd J=9 Hz andJ=5 Hz), 7.16 (1H, d, J=7 Hz), 7.48-7.83 (6H, m), 7.91-8.03 (3H, m),8.31 (1H, d, J=9 Hz), 8.38 (1H, d, J=2 Hz).

Example 2N-[1-[(1-amino-6-isoquinolinyl)methyl]-2-oxo-2-(1-piperidinyl)ethyl]-2-[(2-naphthalenyl-sulfonyl)amino]acetamidehydrochloride (2b)

[0204] 2a.3-(1-amino-6-isoquinolinyl)-2-[[2-[(2-naphthalenylsulfonyl)amino]acetyl]amino]propionicacid hydrochloride 0.16 mL of aqueous 2N NaOH was added to a solution of47 mg of 1k in 1.0 mL of dioxane and 0.5 mL water. After stirring atroom temperature for one hour, the reaction mixture was made acidic (pH2) and extracted with a mixture of n-butanol and dichloromethane.Evaporation of the organic solvents yielded 47 mg of acid 2a. ¹H—NMR 200MHz (CDCl₃/CD₃ OD=3/1) δ: 3.20-3.65 (4H, m), 4.87 (1H, dd, J=7 Hz andJ=5 Hz), 7.10 (1H, d, J=7 Hz), 7.41 (1H, d, J=7 Hz), 7,58-8.03 (8H, m),8.29 (1H, d, 7 Hz), 8.41 (1H, d, 2 Hz)

[0205] 2b.N-[1-[(1-amino-6-isoquinolinyl)methyl]-2-oxo-2-(1-piperidinyl)ethyl]-2-[(2-naphthalenyl-sulfonyl)amino]acetamidehydrochloride

[0206] 47 mg of 2a was suspended in 3 mL of N,N-dimethylformamide andconcentrated in vacuo. The residue was dissolved in 3 mL ofN,N-dimethylformamide and 0.11 mL of piperidine and 33 mg of2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroboratewas added. When the pH of the reaction mixture was not 8-9, the pH wasadjusted to 8-9 with N,N-diisopropyl-ethylamine. After stirring 16 hoursat room temperature the reaction mixture was concentrated in vacuo andchromatographed on silica gel (ethyl acetate/pyridine/acetic acid/water:81/31/18/7 v/v/v/v), The product was dissolved in dichloromethane,washed with water (adjusted to pH 8-9), dried (magnesium sulfate) andconcentrated to give 43 mg of free base. The free base was dissolved ina t-butanol/water mixture, one equivalent hydrochloric acid was addedand lyophilisation gave the title compound 2b. ¹H—NMR 200 MHz (CD₃OD) δ:1.20-1.65 (6H, m), 2.92-3.52 (6H, m), 3.54 (2H, s), 5.12 (1H, t, J=7Hz), 7.17 (1H, d, J=7 Hz), 7.51-8.08 (9H, m), 8.31 (1H, d, J=9 Hz), 8.41(1H, d, J=2 Hz).

Example 33-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[2-[(2-naphthalenylsulfonyl)amino]acetyl]amino]-propionicacid methyl ester hydrochloride (3i)

[0207] 3a. 4-Phenoxythieno[3,2c]pyridine

[0208] Compound 3a was prepared from 4-chlorothieno[3,2c]pyridine (New,J. S. et al., J. Med. Chem. 32, 1147-1156, (1989)) using the proceduredescribed for, 1c. ¹H—NMR 200 MHz (CDCl₃) δ: 7.18-7.28 (3H, m),7.38-7.50 (4H, m), 7.64 (1H, d, J=6 Hz), 8.97 (1H, d, J=6 Hz).

[0209] 3b. 4-Aminothieno[3,2c]pyridine

[0210] The procedure described for 1d was used to prepare 3b from 3a butthe reaction was performed at 155° C. for 3 days. ¹H—NMR 200 MHz (CD₃OD)δ; 7.16 (1H, d, J=6 Hz), 7.48 (1H, d, J=6 Hz), 7.55 (1H, d, J=6 Hz),7.71 (1H, d, J=6 Hz).

[0211] 3c. N-(thieno[3,2c]pyridin-4-yl)benzamide

[0212] The procedure described for 1e was used to prepare 3c from 3b.¹H—NMR 200 MHz (CDCl₃) δ: 7.42-7.77 (6H, m), 7.97-8.22 (3H, m),9.12-9.22 (1H, m).

[0213] 3d. N-(2-formylthieno[3,2c]pyridin-4-yl)benzamide

[0214] 5.2 mL of nBuLi (1.6 N in hexane) was added to a stirred solutionof 0.96 mL of diisopropylamine in 8 mL of tetrahydrofuran under anitrogen atmosphere at −25° C. After stirring for 20 min the solutionwas cooled to −78° C., a solution of 0.88 g of 3c in 14 mL oftetrahydrofuran was added dropwise and the reaction mixture was stirredfor 45 min. Then a mixture of 0.6 mL of N,N-dimethylformamide and 7 mLof tetrahydrofuran was added. The cooling bath was removed, the reactionmixture was allowed to come to room temperature and was poured into anice cold aqueous hydrochloric acid solution (20 mL, 0.5 N). The mixturewas adjusted to pH 7 and the organic solvents were evaporated. Theprecipitate formed was isolated by filtration and dried. The yield was0.98 g of aldehyde 3d. ¹H—NMR 200 MHz (CDCl₃) δ: 7.48 -7.80 (5H, m),8.02-8.14 (2H, m), 8.30-8.38 (2H, m), 10.11 (1H, s).

[0215] 3e. N-[2-(hydroxymethyl)thieno[3,2c]pyridin-4-yl]benzamide

[0216] This compound was prepared from 3d using the procedure describedfor 1g. ¹H—NMR 200 MHz (CDCl₃) δ: 4.88 (2H, s), 7.28 (1H, br.s).7.48-7.76 (5H, m), 7.99-8.22 (3H, m).

[0217] 3f.[[4-(benzoylamino)thieno[3,2c]pyridin-2-yl]methyl][[(1,1-dimethylethoxy)carbonyl]amino]-propanedioicacid diethyl ester

[0218] This compound was prepared from 3e using the procedure describedfor 1h. ¹H—NMR 200 MHz (CDCl₃) δ: 1.28 (6H, t), 1.46 (9H, s), 3.98 (2H,s), 4.18-4.38 (4H, m), 7.05-8.17 (7H, m), 8.75 (1H, br.s).

[0219] 3g. 2-Amino-3-(4-aminothieno[3,2c]pyridin-2-yl)propionic aciddihydrochloride

[0220] This compound was prepared from 3f using the procedure describedfor 1i. ¹H—NMR 200 MHz (D₂O) δ: 3.57-3.80 (2H, m), 4.44 (1H, dd, J=6 Hzand J=7 Hz), 7.45 (1H, d, J=7 Hz), 7.66 (1H, d, J=7 Hz), 7.85 (1H, s).

[0221] 3h. 2-Amino-3-(4-aminothieno[3,2c]pyridin-2-yl)propionic acidmethyl ester dihydrochloride

[0222] This compound was prepared from 3g using the procedure describedfor 1j. ¹H—NMR 200 MHz (CD₃OD) δ: 3.65-3.72 (2H, m), 3.90 (3H, s), 4.53(1H, t, J=6 Hz), 7.47 (1H, dd, J=7 Hz and J=2 Hz), 7.66 (1H, d, J=7 Hz),7.82 (1H, br.s).

[0223] 3i.3-(4-Aminothieno[3,2c]pyridin-2-yl)-2-[[2-[(2-naphthalenylsulfonyl)amino]acetyl]amino]-propionicacid methyl ester hydrochloride

[0224] This compound was prepared from 3h using the procedure describedfor 1k. ¹H—NMR 360 MHz (CD₃OD) δ: 3.32-3.56 (2H, m), 3.61 (2H, s), 3.73(3H, s), 4.75 (1H, dd, J=8 Hz and J=5 Hz), 7.37 (1H, dd, J=1 Hz and J=7Hz), 7.58-7.70 (4H, m), 7.83 (1H, dd, J=8 Hz and J=2 Hz), 7.95-8.05 (3H,m), 8.42 (1H, d, J=2 Hz).

Example 4N-[1-[(4-Aminothieno[3,2c]pyridin-2-yl)methyl]-2-oxo-2-(1-piperidinyl)ethyl]-2-[(2-naphthalenylsulfonyl)amino]acetamidehydrochloride

[0225] 0.06 mL of aqueous 2N NaOH was added to a solution of 20 mg of 3iin 0.25 mL of tetrahydrofuran, 0.1 mL of methanol and 0.25 mL of water.After stirring at room temperature for one hour the reaction mixture wasneutralized and concentrated under reduced pressure. Tituration of theresidue with a small amount of water, acidified to pH 2 withhydrochloric acid, gave3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[2-[(2-naphthalenylsulfonyl)amino]acetyl]amino]-propionicacid hydrochloride. This acid was coupled with piperidine using theprocedure described for 2b. Purification on silica gel(dichloromethane/methanol: 9/1), addition of one equivalent hydrochloricacid, and lyophilisation (t-butanol/water) gave the title compound 4.

[0226]¹H—NMR 360 MHz (CD₃OD) δ: 1.32-1.64 (6H, m),3.10-3.53 (6H, m),3.61 and 3.64 (2H, ABq, J=17 Hz), 5.08 (1H, t, J=6 Hz), 7.35 (1H, dd,J=1 Hz and J=7 Hz), 7.56-7.70 (4H, m), 7.84 (1H, dd, J=9 Hz and J=2 Hz),7.96-8.06 (3H, m), 8.43 (1H, d, J=2 Hz).

Example 5

[0227]N-[1-[(1-Amino-6-isoquinolinyl)methyl]-2-oxo-2-(1-piperidinyl)ethyl]-2-[[(3,4-dihydro-2,2,5,7,8-pentamethyl-2H-1-benzopyran-6-yl)sulfonyl]amino]acetamidehydrochloride (5c)

[0228] 5a. 1,1-Dimethylethyl1-[(1-amino-6-isoquinolinyl)methyl]-2-oxo-2-(1-piperidinyl)ethyl-carbamate

[0229] 144 mg of di-t-butyl dicarbonate in 1 mL of methanol was added toa solution of 100 mg of amino acid 1i in 4.5 mL of methanol and 0.5 mLof triethylamine. After stirring for 10 min the mixture was concentratedunder reduced pressure and coevaporated with toluene. Addition ofmethanol gave a precipitate which was collected by filtration. Theresidue (132 mg) was suspended in N,N-dimethylformamide and 0.04 mL ofpiperidine, 133 mg of2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroboratewas added. After stirring for one hour the reaction mixture wasconcentrated under reduced pressure. Purification by columnchromatography (silica gel, dichloromethane/methanol: 9/1) gave 77 mg of5a. ¹H—NMR 200 MHz (CDCl₃) δ: 0.93-1.59 (15H, m), 2.95-3.55 (6H, m),4.93, (1H, t, J=7 Hz), 6.95 (1H, dd J=6 Hz and J=2 Hz), 7.37 (1H, dd,J=8 Hz and J=2 Hz), 7.41 (1H, d, J=2 Hz), 7,78-7.84 (2H, m).

[0230] 5b.[(3,4-dihydro-2,2,5,7,8-pentamethyl-2H-1-benzopyran-6-yl)sulfonyl]amino]aceticacid

[0231] 0.33 g of glycine was dissolved in 8.8 mL of aqueous 1N sodiumhydroxide, a solution of 1.2 g of(3,4-dihydro-2,2,5,7,8-pentamethyl-2H-1-benzopyran-6-yl)sulfonylchloridein 4 mL dioxane and 4 mL ether was added and stirred for 16 h.The-aqueous solution was acidified to pH 3 with hydrochloric acid andextracted twice with ether. The combined organic layers were dried overmagnesium sulfate and concentrated to give 1.15 g of 5b. ¹H—NMR 200 MHz(CDCl₃) δ: 1.32 (6H, s), 1.82 (2H, t, J=7 Hz), 2.12 (3H, s ), 2.53 (3H,s), 2.55 (3H, s), 2.64 (2H, t, J=7 Hz), 3.77 (2H, s).

[0232] 5c.N-[1-[(1-Amino-6-isoquinolinyl)methyl]-2-oxo-2-(1-piperidinyl)ethyl]-2-[[(3,4-dihydro-2,2,5,7,8-pentamethyl-2H-1-benzopyran-6-yl)sulfonyl]amino]acetamidehydrochloride

[0233] To 70 mg of 5a in 2 mL of dichloromethane was added 0.01 mL ofthioanisol and 0.5 mL of trifluoroacetic acid. After stirring for 2hours at room temperature the reaction mixture was concentrated andcoevaporated with water and subsequently with N,N-dimethylformamide. Theresidue and 72 mg of 5b was suspended in 3 mL of N,N-dimethylformamideand N-ethylmorpholine was added until pH 8. The mixture was cooled at 0°C. and 45 mg of 1-hydroxybenzotriazole and 45 mg ofN,N-dicyclohexylcarbodiimide were added. After stirring for 16 hours atroom temperature the mixture was concentrated in vacuo. Dichloromethanewas added to the residue and filtered. The filtrate was purified onsilica gel (dichloromethane/methanol: 95/5) to give the free base. Tothis free base was added one equivalent hydrochloric acid andlyophylisation (t-butanol/water) gave 75 mg of the title compound 5c.¹H—NMR 200 MHz (CDCl₃) δ: 1.32 (6H, s), 1.60-187 (8H, m), 2.13 (3H, s),2.56-3.78 (16H, m), 5.28 -5.38 (1H, m), 7.15-7.21 (2H, m), 7.59 (1H, d,J=7 Hz), 7.75-7.86 (2H, m).

Example 6N-[1-[(1-Amino-6-isoquinolinyl)methyl]-2-oxo-2-(1-piperidinyl]ethyl]-2-[[(4-methylphenyl)sulfonyl]amino]acetamidehydrochloride

[0234] The procedure described for 5c was used. Deprotection of 5a andsubsequently coupling with [[(4-methylphenyl)sulfonyl]amino]acetic acid(McChesney, E. W. and Swann, W. K., J. Am. Chem. Soc. 59, 1116 (1937))yielded the title compound 6. ¹H—NMR 200 MHz (CD₃OD) δ: 1.22-1.68 (6H,m), 2.42 (3H, s), 3.03-3.68 (8H, m), 5.21 (1H, dd, J=6 Hz and J=7 Hz),7.19 (1H, d, J=7 Hz), 7.35-7.80 (7H, m), 8.32 (1H, d, J=9 Hz).

Example 7(2S)-N-[1-[(1-amino-6-isoquinolinyl)methyl]-2-oxo-2-(1-piperidinyl)ethyl]-3-hydroxy-2-[(2-naphthalenylsulfonyl)amino]propanamidehydrochloride

[0235] The procedure described for 5c was used. Deprotection of 5a andcoupling with (2S)-3-hydroxy-2-[(2-naphthalenyisulfonyl)amino]propanoicacid (prepared from L-serine and 2-naphthalenylsulfonylchloride usingthe procedure described for 5b) yielded 7. ¹H—NMR 200 MHz (CD₃OD) δ:1.12-1.58 (6H, m), 2.57-2.69 (1H, m) 2.95-3.95 (8H, m), 4.9-5.1 (1H, m),7.11-7.22 (1H, m), 7.43-8.16 (9H, m), 8.24-8.34 (1H, m), 8.41-8.45 (1H,m).

Example 8(3S)-4-[[1-[(1-amino-6-isoquinolinyl)methyl]-2-oxo-2-(1-piperidinyl)ethyl]amino]-3-[[(4-methoxy-2,3,6-trimethylphenyl)sulfonyl]amino]4-oxo-butanoicacid 1,1-dimethylethylester hydrochloride

[0236] The procedure described for 5c was used. Deprotection of 5a andcoupling with 121 mg of(2S)-2-[[(4-methoxy-2,3,6-trimethylphenyl)sulfonyl]amino]butanedioicacid 4-(1,1-dimethyl-ethyl)ester (prepared from Asp(OtBu)—OH and(4-methoxy-2,3,6-trimethylphenyl)-sulfonylchloride using the proceduredescribed for 5b) yielded after purification 8 as a mixture ofdiastereomers (1:1). ¹H—NMR 200 MHz (CD₃OD) δ: 1.30 and 1.35 (9H, 2× s),1.38-1.70 (6H, m), 2.12 and 2.14 (3H, 2× s) 2.18-2.51 (2H, m), 2.53 and2.54, (3H, 2× s), 2.63 (3H, s) 2.85-3.54 (6H, m), 3.81 and 3.86 (3H, 2×s), 3.95-4.11 (1H, m), 5.03-5.21 (1H, 6.73 and 6.75 (1H, 2× s),7.17-7.24 (1H, m), 7.52-7.78 (3H, m), 8.30-8.38 (1H, m).

Example 9(3S)-4-[[1-[(1-amino-6-isoquinolinyl)methyl]-2-oxo-2-(1-piperidinyl)ethyl]amino]-3-[[(4-methoxy-2,3,6-trimethylphenyl)sulfonyl]amino]4-oxo-butanoicacid hydrochloride

[0237] 10 mL of aqueous 1N hydrochloric acid was added to 45 mg of 8.After stirring for 6 h at 50° C. the reaction mixture was concentratedin vacuo. The residue was purified (silica gel, ethyl acetate/pyridine/acetic acid/water: 81/31/18/7 and LH20,methanol/dichloromethane: 1/1) to yield the free base. Addition of oneequivalent hydrogen chloride and lyophilisation (t-butanol/water)yielded 32 mg of 9 as a mixture of diastereoisomers. ¹H—NMR 200 MHz(CD₃OD) δ: 1.3-1.7 (6H, m), 2.14 (3H, s) 2.20-2.38 (1H, m), 2.53 and2.55, (3H, 2× s), 2.62 and 2.64 (3H, 2× s), 2.71-3.98 (8H, m), 3.85 and3.87 (3H, 2× s), 5.08-5.21 (1H, m), 6.61 (1H br.s.), 7.07 (1H, d, J=7Hz), 7.42-7.64 (3H, m), 8.21-8.28 (1H, m).

Example 103,4-dihydro-2,2,5,7,8-pentamethyl-2H-1-benzopyran-6-ylsulfonic acid2-[[[1-[(1-amino-6-isoquinolinyl)methyl]-2-oxo-2-(1-pineridinyl)ethyl]amino]carbonyl]hydrazidehydrochloride

[0238] To 100 mg of 5a in 2 mL of dichloromethane was added 2 mL oftrifluoroacetic acid. After stirring for 15 minutes at room temperaturethe reaction mixture was concentrated and coevaporated with toluene. Theresidue was suspended in 3 mL N,N-dimethylformamide and 0.14 mL ofN,N-diisopropylethylamine and 76 mg of Boc Azagly 4-nitrophenylester(Gante, J. And Weitzel., R., Liebigs Ann. Chem. 349 (1990)) were added.After stirring for 2 hours at room temperature the reaction mixture wasconcentrated under reduced pressure and crystallized fromdiethylether/dichloromethane to give 89 mg of the coupled product. To 50mg of this product were added 1.8 mL of acetic acid and 0.2 mL ofaqueous 1N hydrochloric acid. After stirring for 45 minutes at roomtemperature the reaction mixture was concentrated under reduced pressureand coevaporated with toluene. The residue was suspended in 2 mL ofN,N-dimethylformamide and 0.06 mL of N,N-diisopropylethylamine and asolution of 38 mg of(3,4-dihydro-2,2,5,7,8-pentamethyl-2H-1-benzopyran-6-yl)sulfonylchloridein 1 mL of N,N-dimethylformamide was added. After stirring for 3 hoursat room temperature the reaction mixture was concentrated under reducedpressure and purification yielded 38 mg of 10. ¹H—NMR 200 MHz (CD₃OD) δ:0.95-1.65 (6H, m), 1.24 (3H, s), 1.28 (3H, s), 1.80 (2H, t, J=7 Hz),2.33 (3H, s), 2.55 (3H, s), 2.64-3.48 (8H, m), 5.00 (1H, t, J=7 Hz)),7.10 (1H, d, J=7 Hz), 7.44 (1H, dd, J=2 Hz and J=9 Hz), 7.59 (1H, d, J=7Hz), 7.64 (1H, d, J=2 Hz), 8.22 (1H, d, J=9 Hz).

Example 111-[3(1-amino-6-isoquinolinyl)-2-[(2-naphthalenylsulfonyl)amino]-1-oxopropyl]-4-methylpiperidinehydrochloride (11b)

[0239] 11a.3-(1-amino-6-isoquinolinyl)-2-[(2-naphthalenylsulfonyl)amino]propionicacid methyl ester hydrochloride

[0240] 0.10 g of 2-naphthalenylsulfonylchloride dissolved in 0.8 mL ofdichloromethane and 0.2 mL of dioxane was added to a solution of 0.13 gof 1j in 6 mL dichloromethane and 0.17 mL of triethylamine at 0° C.After stirring at room temperature for 1 hour water was added , sodiumhydroxide was added until pH 8-9 and the mixture was extracted withdichloromethane. The dichloromethane extract was dried (MgSO₄) andconcentrated. Purification (silica gel, dichloromethane/methanol 95/5),addition of one equivalent of hydrochloric acid and lyophilisation(t-butanol/water) gave 69 mg of 11a. ¹H—NMR 200 MHz (CDCl₃) δ: 3.00-3.34(2H, m), 3.49 (3H, s), 4.34 (1H, dd, J=5 Hz and J=9 Hz), 6.72 (1H, d,J=7 Hz), 7.25-7.87 (9H, m), 8.04 (1H, d, J=8 Hz), 8.16 (1H, d, J=2 Hz).

[0241] 11b.1-[3-(1-amino-6-isoquinolinyl)-2-[(2-naphthalenylsulfonyl)amino]-1-oxopropyl]-4-methylpiperidinehydrochloride

[0242] Compound 11a was saponified and subsequently coupled with4-methylpiperidine using the procedure described for 4 to give 11b.¹H—NMR 200 MHz (CD₃OD) δ: 0.15-3.24 (12H, m), 3.75-4.24 (2H, m),4.57-4.72 (1H, m), 6.71-7.91 (10H, m), 8.11-8.19 (2H, m)

Example 121-[3-(1-amino-6-isoquinolinyl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]amino]-1-oxopropyl]-4-methylpiperidinehydrochloride (12b)

[0243] 12a. (7-methoxy-2-naphthalenyl)sulfonyl chloride

[0244] 2-(7-hydroxynaphthalenyl)sulfonic acid was methylated (J.Org.Chem. 57, 2631 (1992)) and subsequently treated with thionyl chloride(Hel. Chim. Acta 176, 1653 (1959) using the procedures described in theindicated literature to give the title compound 12a. M.p.: 81-85° C.

[0245] 12b.1-[3(1amino-6-isoquinolinyl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]amino]-1-oxopropyl]-4-methylpiperidinehydrochloride

[0246] Using the procedure described for 11a, the reaction of methylester 1j and sulfonyl chloride 12a gave3-(1-amino-6-isoquinolinyl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl)amino]propionicacid methyl ester. This ester was saponified and subsequently coupledwith 4-methylpiperidine using the procedure described for the previousexample to give the title compound 12b. ¹H—NMR 200 MHz (CD₃OD) δ:0.18-0.85 (5H, m), 1.22-1.58 (3H, m), 1.78-3.18 (4H, m), 3.80-4.26 (2H,m), 3.88 (3H, s), 4.57-4.72 (1H, m), 6.97-7.04 (1H, m), 7.20-7.79 (8H,m) 8.01-8.14 (2H, m).

Example 131-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[(2-naphthalenylsulfonyl)amino]-1-oxopropyl]-4-methylpiperidinehydrochloride

[0247] Using the procedure described for 11a, the reaction of compound3h and 2-naphthalenylsulfonylchloride gave3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[(2-naphthalenyl-sulfonyl)amino]propionicacid methyl ester hydrochloride. This compound was saponified andcoupled with 4-methylpiperidine according to the procedure described for11b giving the title compound 13. ¹H—NMR 200 MHz (CD₃OD) δ: 0.1-3.3(12H, m), 3.72-4.18 (2H, m), 4.54-4.68 (1H, m), 7.16-7.23 (1H, m),7.38-8.01 (8H, m), 8.27-8.32 (1H, m).

Example 141-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]amino]-1-oxopropyl]-4-(methylsulfonyl)piperazinehydrochloride (14c)

[0248] 14a.3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]-amino]propionicacid hydrochloride

[0249] The procedure described for 11a was used to prepare3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]amino]propionic acid methyl ester from 3h and 12a. This sulfonamide (130 mg)was dissolved in 3 mL of dioxane and 1.1 mL of water and 0.4 mL ofaqueous 2N NaOH was added. After stirring at room temperature for 2hours the reaction mixture was neutralized and concentrated underreduced pressure. The residue was subsequently titurated withdichloromethane and with a small amount of water acidified to pH 3 withhydrochloric acid to give 114 mg of compound 14a. TLC (silica gel, ethylacetate/pyridine/acetic acid/water: 81/31/18/7) rf=0.29.

[0250] 14b. 1-Methylsulfonylpiperazine

[0251] 3.5 mL of methane sulfonylchloride in 50 mL of dichloromethanewas added slowly to a solution of 4.9 g of 1-formylpiperazine and 6.1 mLof triethylamine in 100 mL of dichloromethane at 0° C. After stirringfor 1 hour water was added and the organic layer was separated. Theaqueous layer was extracted several times with dichloromethane. Thecombined organic layers were dried (MgSO₄) and concentrated to yield 5.8g of 1-formyl-4-(methylsulfonyl)piperazine. The crude product wasdissolved in 15 ml of ethanol and 15 mL of aqueous 2N NaOH and stirredfor 1.5 hours at 80° C. After cooling to room temperature water wasadded and extracted several times with dichloromethane. The combineorganic layers were dried (MgSO₄) and concentrated to give 2.9 g of thetitle compound 14b. ¹H—NMR 200 MHz (DMSOd6) δ: 2.70-2.77 (4H, m), 2.83(3H, s), 2.95-3.02 (4H, m).

[0252] 14c.1-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]-1-oxopropyl]-4-(methylsulfonyl)piperazinehydrochloride

[0253] Using the procedure described for 2b, 14a was coupled with 14b togive the title compound 14c.

[0254]¹H—NMR 400 MHz (CD₃OD) δ: 2.66 (3H, s), 2.62-3.69 (10H, m), 3.97(3H, s) 4.6-4.7 (1H, m), 7.14 (1H, d, J=7 Hz), 7.29-7.35 (3H, m),7.46-7.60 (2H, m), 7.81-7.86 (2H, m), 8.18 (1H, bs.s).

Example 151-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]amino]-1-oxopropyl]morpholinehydrochloride

[0255] Using the procedure described for 2b, 14a was coupled withmorpholine to give 15. ¹H—NMR 200 MHz (CD₃OD) δ: 3.07-3.52 (10H, m),3.94 (3H, s), 4.62 (1H, dd, J=9 Hz and J=5 Hz), 7.19-7.58 (6H, m),7.76-7.82 (2H, m), 8.13 (1H, d, J=2 Hz).

Example 161-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[[(5-dimethylamino)naphthalenyl]sulfonyl]-amino]-1-oxopropyl]-4-methylpiperidinehydrochloride (16b)

[0256] 16a.3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[[(5-dimethylamino)naphthalenyl]sulfonyl]-amino]-propionicacid hydrochloride

[0257] Using the procedure described for 14a, coupling of 3h and(5-dimethylaminonaphthalenyl)-sulfonyl chloride gave title compound 16aTLC (silica gel, ethyl acetate/pyridine/acetic acid/water: 81/31/18/7)rf=0.3.

[0258] 16b.1-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[[(5-dimethylamino)naphthalenyl]sulfonyl]-amino]-1-oxopropyl]-4-methylpiperidinehydrochloride

[0259] Using the procedure described for 2b, 16a was coupled with4-methylpiperidine to give the title compound 16b. ¹H—NMR 200 MHz(CD₃OD) δ: 0.1-3.3 (12H, m), 2.82 (3H, s), 3.53-4.28 (2H, m), 4.41-4.52(1H, m), 7.02-7.58 (6H, m), 8.12-8.24 (2H, m), 8.40-8.51 (1H, m)

Example 171-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[[(5-dimethylamino)naphthalenyl]sulfonyl]-amino]-1-oxopropyl]-4-(methylsulfonyl)piperazinehydrochloride

[0260] Using the procedure described for 2b, 16a was coupled with1-methylsulfonylpiperazine to give the title compound 17. ¹H—NMR 200 MHz(CD₃OD) δ: 2.64 (3H, s), 2.83 (6H, s), 2.76-3.46 (10H, m), 4.50 (1H, dd,J=8 Hz and J=7 Hz), ), 7.03-7.15 (2H, m), 7.25 (1H, br.s), 7.43-7.57(3H, m), 8.14-8.20 (2H, m), 8.41-8.48 (1H, m)

Example 181-[1-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]amino]-1-oxopropyl]-4-methylpiperidinehydrochloride

[0261] Using the procedure described for 2b, 14a was coupled with4-methylpiperidine to give the title compound 18. ¹H—NMR 200 MHz (CD₃OD)δ: 0.1-3.3 (12H, m), 3.93 (3H, s), 3.65-4.17 (2H, m), 4.52-4.65 (1H, m),7.09-7.87 (8H, m), 8.18-8.23 (1H, m).

Example 191-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]amino]-1-oxopropyl]-4-ethylpiperidinehydrochloride

[0262] Using the procedure described for 2b, 14a was coupled with4-ethylpiperidine to give the title compound 19. ¹H—NMR 200 MHz (CD₃OD)δ: 0.1-3.3 (14H, m), 3.72-3.83 (1H, m), 3.93 and 3.94 (3H, 2× s),3.94-4.16 (1H, m), 4.52-4.63 (1H, m), 7.12-7.61 (6H, m) 7.76-7.85 (2H,m), 8.17-8.24 (1H, m).

Example 201-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[[(5-dimethylamino)naphthalenyl]sulfonyl]-amino]-1-oxopropyl]morpholinehydrochloride

[0263] Using the procedure described for 2b, 16a was coupled withmorpholine to give the title compound 20. ¹H—NMR 200 MHz (CD₃OD) δ: 2.82(6H, s), 2.99-3.46 (10H, m), 4.45 (1H, dd, J=8 Hz and J=7 Hz), ),7.01-7.13 (2H, m), 7.24 (1H, br.s), 7.41-7.57 (3H, m) 8.14-8.19 (2H, m),8.40-8.47 (1H, m)

Example 211-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]amino]-1-oxopropyl]-4-formylpiperazinehydrochloride

[0264] Using the procedure described for 2b, 14a was coupled with1-formylpiperazine to give the title compound 21. ¹H—NMR 400 MHz (CD₃OD)δ: 2.81-3.68 (10H, m), 3.72 (3H, s), 4.66 (1H, dd, J=6 Hz and J=8 Hz)),7.08-7.10 (1H, m), 7.25-7.32 (3H, m), 7.49 (1H, d, J=7 Hz), 7.58 (1H,dd, J=2 Hz and J=9 Hz), 7.77-7.81 (2H, m), 7.92 and 7.98 (1H, 2× s),8.17 (1H, bs.s).

Example 221-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]amino]-1-oxopropyl]-4-methoxypiperidinehydrochloride (22b)

[0265] 22a. 4-Methoxypiperidine hydrochloride

[0266] 5.8 g of di-t-butyl dicarbonate was added to a solution of 3.13 gof 4-hydroxypiperidine in 29 mL of tetrahydrofuran and 7 mL of pyridine.After 16 hours at room temperature the mixture was concentrated andcoevaporated with toluene. The residue was dissolved in 23 mL oftetrahydrofuran and 3.3 mL of methyl iodide was added. To this solution1.2 g of sodium hydride (60% dispersion) was added in small potions.After stirring for 2 hours at room temperature methanol was addedcarefully to destroy the excess sodium hydride. The reaction mixture wasconcentrated in vacuo. Water was added, the pH adjusted to 4 andextracted with ethyl acetate. The ethyl acetate extract was dried(MgSO₄) and concentrated. Column chromatography (silica gel,toluene/ethyl acetate: 4/1) yielded 2.7 g of 1-Boc-4-methoxypiperidine.This compound was cooled at 0° C. and 30 mL of 3N hydrochloric acid inmethanol was added. After stirring for 19 hours at room temperature thereaction mixture was concentrated to give 2.4 g of 4-methoxy-piperidinehydrochloride. ¹H—NMR 200 MHz (CD₃OD) δ: 1.76-2.12 (4H, m), 3.03-3.61(5H, m), 3.36 (3H, s).

[0267] 22b.1-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]amino]-1-oxopropyl]-4-methoxypiperidinehydrochloride

[0268] Using the procedure described for 2b, 14a was coupled with 22a togive the title compound 22b.

[0269]¹H—NMR 400 MHz (CD₃OD) δ: 1.82-1.62 (4H, m), 2.90-3.64 (7H, m),3.21 and 3.22 (3H, 2× s), 3.94 (3H, s), 4.59-4.64 (1H, m), 7.11 (1H, d,J=7 Hz), 7.25-7.34 (3H, m), 7.49-7.58 (2H, m), 7.77-7.82 (2H, m), 8.18(1H, br.s).

Example 231-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]amino-1-oxopropyl]-4-[(trifluormethyl)sulfonyl]piperazinehydrochloride (23b)

[0270] 23a. 1-(trifluormethyl)sulfonyl]piperazine hydrochloride

[0271] 2.0 mL of trifluormethanesulfonic anhydride in 10 mL ofdichloromethane was slowly added to a stirred solution of 2.0 g ofBoc-piperazine and 1.65 mL of triethylamine in 31 mL of dichloromethaneat −78° C. After 1 hour the reaction mixture was allowed to warm to 0°C., poured into water, neutralized and extracted with dichloromethane.The dichloromethane extract was washed with 5% NaHCO₃, dried (MgSO₄) andconcentrated. Column chromatography (silica gel, toluene/ethyl acetate:9/1) yielded 1.6 g of 1-Boc-4-[(trifuormethyl)sulfonyl]piperazine.

[0272] 0.2 g of this compound was cooled at 0° C. and 3 mL of 3Nhydrochloric acid in methanol was added. After stirring for 19 hours atroom temperature the reaction mixture was concentrated to give 0.1 g of1-(trifluormethyl)sulfonyl]piperazine hydrochloride. ¹⁹F—NMR 188 MHz(CD₃OD) δ: −76.6.

[0273] 23b.1-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]amino-1-oxopropyl]-4-[(trifluormethyl)sulfonyl]piperazinehydrochloride

[0274] Using the procedure described for 2b, 14a was coupled with 23a togive the title compound 23b.

[0275]¹⁹F—NMR 188 MHz (CD₃OD) δ: −78.3.

Example 241-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]amino]-1-oxopropyl]-4-methylpiperazinedihydrochloride

[0276] Using the procedure described for 2b, 14a was coupled with1-methylpiperazine to give the title compound 24. ¹H—NMR 400 MHz (CD₃OD)δ: 1.99-2.51 (4H, m), 2.23 (3H, s), 3.12-3.72 (6H, m), 3.94 (3H, s),4.63 (1H, dd, J=5 Hz and J=9 Hz), 7.18 (1H, d, J=7 Hz), 7.26-7.31 (2H,m), 7.39 (1H, s), 7.46 (1H, d, J=7 Hz), 7.56 (1H, dd, J=2 Hz and J=9Hz), 7.78-7.82 (2H, m), 8.15 (1H, d, J=2 Hz).

Example 251-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]amino]-1-oxopropyl]thiomorpholinehydrochloride

[0277] Using the procedure described for 2b, 14a was coupled withthiomorpholine to give the title compound 25. ¹H—NMR 400 MHz (CDCl₃) δ:1.87-2.41 (4H, m), 3.03-3.75 (6H, m), 3.95 (3H, s), 4.56 (1H, dd, J=5 Hzand J=8 Hz)), 6.94 (1H, d, J=7 Hz), 7.21 (1H, d, J=3 Hz), 7.28-7.35 (2H,m), 7.57 (1H, d, J=6 Hz), 7.63 (1H, dd, J=2 Hz and J=9 Hz), 7.76-7.84(2H, m), 8.25 (1H, bs.s).

Example 263-(4-aminothieno[3,2c]pyridin-2-yl)-N-(2-methoxyethyl)-2-[[(7-methoxy-2-naphthalenyl)-sulfonyl]amino]-N-methyl-propanamidehydrochloride

[0278] Using the procedure described for 2b, 14a was coupled withN-methyl-2-methoxyethylamine to give the title compound 26. ¹H—NMR 400MHz (CD₃OD) δ: 2.73 and 2.98 (3H, 2× s), 3.08 and 3.28 (3H, 2× s),2.85-3.48 (5H, m), 3.91 (3H, s), 4.01-4.27 (1H, m), 4.56-4.76 (1H, m),6.89-7.02 (1H, m), 7.16-7.78 (7H, m), 8.04-8.19 (1H, m).

Example 271-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]amino]-1-oxopropyl]-4-(1-oxoethyl)piperidinehydrochloride

[0279] Using the procedure described for 2b, 14a was coupled with4-(1-oxoethyl)piperidine to give the title compound 27. ¹H—NMR 400 MHz(CD₃OD) δ: 0.52-0.72 (1H, m), 0.97-1.18 (1H, m), 1.53-1.71 (2H, m), 1.92and 1.98 (3H, 2× s), 2.04-2.56 (2H, m), 2.76-2.92 (1H, m), 3.08-3.28(2H, m), 3.73-4.05 (2H, m), 3.94 (3H, s), 4.57-4.63 (1H, m), 7.09 (1H,d, J=7 Hz), 7.23-7.35 (3H, m), 7.53-7.60 (2H, m), 7.75-7.86 (2H, m),8.17 and 8.22 (1H, 2× br.s.).

Example 281-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]amino]-1-oxopropyl]-4-methylenepiperidinehydrochloride (28b)

[0280] 28a. 4-methylenepiperidine hydrochloride

[0281] 6 g of di-t-butyldicarbonate, 2 g of 4-piperidone monohydratehydrochloride and 6.3 mL of pyridine were dissolved in 25 mL oftetrahydrofuran. After stirring for 4 days at room temperature thereaction mixture was concentrated, water was added, the pH adjusted to 3and extracted with ethyl acetate. The ethyl acetate extracts were dried(magnesium sulfate) and concentrated. This residue (0.6 g) wastransformed into Boc-4-methylenepiperidine using the procedure describedin J. Am. Chem. Soc. 101, 7032 (1979). Boc-4-methylenepiperidine wasdissolved in 3N hydrochloric acid in methanol and stirred for 19 hoursat room temperature. Evaporation yielded 0.24 g of the title compound28a. ¹H—NMR 200 MHz (CDCl₃) δ: 2.57 (4H, br.s), 3.22 (4H, br.s), 4.88(2H, s).

[0282] 28b.1-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]amino]-1-oxopropyl]-4-methylenepiperidinehydrochloride

[0283] Using the procedure described for 2b, 14a was coupled with 28a togive the title compound 28b.

[0284]¹H—NMR 400 MHz (DMSOd6) δ: 1.55-2.04 (4H, m), 2.89-3.5 (6H, m),3.88 (3H, s), 4.49-4.67 (3H, m), 7.43-7.78 (8H, m), 8.09 (1H, 2× br.s).

Example 292-[[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]amino-1-oxopropyl](cyclopropyl)amino]aceticacid ethyl ester hydrochloride

[0285] The procedure described for 2b was used to couple 14a with[(cyclopropyl)amino]acetic acid ethyl ester giving the title compound29. [(Cyclopropyl)amino]acetic acid ethyl ester was preparedcyclopropylbromide and glycine ethyl ester using the procedure describedby J. T. Suh et al.(J. Med. Chem. 28, 57-66 (1985)). ¹H—NMR 400 MHz(CD₃OD) δ: 0.76-1.12 (4H, m), 1.24 (3H, t, J=7 Hz), 2.93-3.02 (2H, m),3.28-3.36 (1H, m), 3.84 and 4.20 (2H, ABq, J=17 Hz), 3.93 (3H, s), 4.14(2H, q, J=7 Hz), 5.10 (1H, dd, J=4 Hz and J=10 Hz), 6.86 (1H, d, J =6Hz), 7.15-7.70 (7H, m), 8.05 (1H, s),

Example 301-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[[1-(1R,4S)-(7,7-dimethyl-2-oxobicyclo[2,2,1]hept-1-yl)methyl]sulfonyl]amino]-1-oxopropyl]-4-methylpiperidinehydrochloride (30b)

[0286] 30a. 1,1-Dimethylethyl1-[(4-aminothieno[3,2c]pyridin-2-yl)methyl]-2-oxo-2-[1-4-methyl-piperidinyl)]ethylcarbamate

[0287] The procedure described for 5a was used. Amino acid 3g wasprotected with the t-butyloxy carbamate group and subsequently coupledwith 4-methylpiperidine to give the title compound 30a. ¹H—NMR 200 MHz(CDCl₃) δ: 0.1-3.4 (12H, m), 1.40 (9H, s), 3.78-3.95 (1H, m), 4.42-4.56(1H, m), 4.87-4.97 (1H, m), 7.04-7.11 (2H, m), 7.78 (1H, d, J=7 Hz).

[0288] 30b.1-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[[1-(1R,4S)-(7,7-dimethyl-2-oxobicyclo[2.2.1]hept-1-yl)methyl]sulfonyl]amino]-1-oxopropyl]-4-methylpiperidinehydrochloride

[0289] To 82 mg of 30a in 2.2 mL of dichloromethane was added 2.2 mL oftrifluoroacetic acid. After stirring for 1 hour at room temperature thereaction mixture was concentrated and coevaporated with toluene. Theresidue was dissolved in 3 mL of dichloromethane and 0.12 mL oftriethylamine, cooled at 0° C. and 55 mg of(−)camphor-10-sulfonylchloride was added. After stirring for 16 hours atroom temperature water was added, the pH was adjusted to 8-9 and themixture was extracted with dichloromethane. The extract was dried(magnesium sulfate) and concentrated. Purification on silica gel(dichloromethane/methanol: 9/1) afforded the free base. Addition of oneequivalent hydrochloric acid and lyophilisation gave 58 mg of the titlecompound 30b. ¹H—NMR 200 MHz (CD₃OD) δ: 0.1-3.5 (28H, m), 3.93-4.16 (1H,m), 4.37-4.58 (1H, m), 7.39-7.48 (1H, m), 7.58-7.73 (2H, m).

Example 311-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(3,4-dihydro-2,2,5,7,8-pentamethyl-2H-1-benzopyran-6-yl)sulfonyl]amino]-1-oxopropyl]-4-methylpiperidinehydrochloride

[0290] The procedure described for 30b was used to couple 30a with(3,4-dihydro-2,2,5,7,8-pentamethyl-2H-1-benzopyran-6-yl)sulfonylchloride giving the title compound 31. ¹H—NMR 200 MHz (CD₃OD) δ: 0.1-1.6(14H, m), 1.82 (2H, t, J=7 Hz), 2.02-3.3 (15H, m), 3.45-3.75 (1H, m),4.11-4.48 (2H, m), 7.25-7.32 (1H, m), 7.44-7.63 (2H, m).

Example 321-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(2-dibenzofuranyl)sulfonyl]amino]-1-oxopropyl]-4-methylpiperidinehydrochloride

[0291] The procedure described for 30b was used to couple 30a with(2-dibenzofuranyl)sulfonyl chloride giving the title compound 32. ¹H—NMR200 MHz (CD₃OD) δ: 0.1-0.75 (5H, m), 1.25-1.56 (3H, m), 1.92-2.50 (1H,m), 2.71-3.3 (3H, m), 3.75-4.23 (2H, m), 4.54-4.69 (1H, m), 6.99-7.05(1H, m), 7.29-7.48 (6H, m), 7.86-8.09 (2H, m), 8.37-8.42 (1H, m).

Example 331-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[2-[5-(2-pyridinyl)thienyl]sulfonyl]amino]-1-oxopropyl]-4-methylpiperidinehydrochloride

[0292] The procedure described for 30b was used to couple 30a with[5-(2-pyridinyl)thienyl]sulfonyl chloride giving the title compound 33.¹H—NMR 200 MHz (CD₃OD) δ: 0.05-1.69 (8H, m), 2.25-2.60 (1H, m),2.85-3.41 (3H, m), 3.78-4.00 (1H, m), 4.17-4.37 (1H, m), 4.59-4.72 (1H,m), 7.13-7.22 (1H, m), 7.29-7.60 (5H, m), 7.82-7.88 (2H, m), 8.47-8.55(1H, m).

Example 341-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(6,7-dimethoxy-2-naphthalenyl)sulfonyl]-amino]-1-oxopropyl]-4-methylpiperidinehydrochloride (34b)

[0293] 34a. 2-(6,7-Dimethoxynaphthalenyl)sulfonyl chloride

[0294] 2-(6,7-dihydroxynaphthalenyl)sulfonic acid was methylated (J.Org. Chem. 57, 2631 (1992)) and subsequently treated with thionylchloride (Hel. Chim. Acta 176, 1653 (1959) using the proceduresdescribed in the indicated literature to give the title compound 34a.M.p.: 111-115° C.

[0295] 34b.1-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(6,7-dimethoxy-2-naphthalenyl)sulfonyl]amino]-1-oxopropyl]-4-methylpiperidinehydrochloride

[0296] The procedure described for 30b was used to couple 30a with 34agiving the title compound 34b. ¹H—NMR 200 MHz (CD₃OD) δ: 0.05-1.58 (8H,m), 1.88-3.3 (4H, m), 3.72-4.19 (2H, m), 3.97 (3H, s), 3.98 and 3.99(3H, 2× s), 4.48-4.64 (1H, m), 7.26-7.32 (3H, m), 7.46-7.79 (4H, m),8.12 (1H, d, J=2 Hz).

Example 351-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[2-[5-(3-isoxazolyl)thienyl]sulfonyl]amino]-1-oxopropyl]-4-methylpiperidinehydrochloride

[0297] The procedure described for 30b was used to couple 30a with[5-(3-isoxazolyl)thienyl]sulfonyl chloride giving the title compound 35.¹H—NMR 200 MHz (CD₃OD) δ: 0.05-1.75 (8H, m), 2.25-2.64 (1H, m),2.84-3.40 (3H, m), 3.79-4.02 (1H, m), 4.18-4.39 (1H, m), 4.61-4.74 (1H,m), 6.75-6.79 (1H, m), 7.14-7.20 (1H, m), 7.38-7.61 (4H, m), 8.46 (1H,d, J=2 Hz).

Example 361-[3-(4-aminothieno[3,2c]pyridin-2-yl-2-[[(4,6-dimethoxy-2naphthalenyl)sulfonyl]-amino]-1-oxopropyl]-4-methylpiperidinehydrochloride (36b)

[0298] 36a. (4,6-Dimethoxy-2-naphthalenyl)sulfonyl chloride

[0299] (4,6-dihydroxy-2-naphthalenyl)sulfonic acid was methylated (J.Org. Chem. 57, 2631 (1992)) and subsequently treated with phosphorousoxychloride (J. Am. Chem. Soc. 74, 2006 (1952)) using the proceduresdescribed in the indicated literature to give the title compound 36a.M.p.: 133.9-134.5° C.

[0300] 36b.1-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(4,6-dimethoxy-2-naphthalenyl)sulfonyl]amino]-1-oxopropyl]-4-methylpiperidinehydrochloride

[0301] The procedure described for 30b was used to couple 30a with 36agiving the title compound 36b. ¹H—NMR 200 MHz (CD₃OD) δ: 0.0-3.3 (12H,m), 3.60-3.76 (1H, m), 3.93 and 3.94 (3H, 2× s), 4.04 (3H, s), 3.97-4.18(1H, m), 4.44-4.60 (1H, m), 7.04-7.57 (6H, m), 7.78-7.87 (2H, m).

Example 371-[3-(1-Amino-7-isoquinolinyl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]amino]-1-oxopropyl]-4-methylpiperidinehydrochloride (37j)

[0302] 37a. 7-Bromoisoquinoline N-oxide hydrochloride

[0303] Compound 37a was prepared from 7-bromoisoquinoline (Tyson, F. L.,J. Am. Chem. Soc. 61, 183 (1939), this procedure gave a mixture of7-bromoisoquinoline and 5-bromoisoquinoline) using the proceduredescribed for 1a. The title compound was contaminated with the5-bromo-isoquinoline N-oxide hydrochloride. M.p. 107.0-112.5° C.

[0304] 37b. 7-Bromo-1-chloroisoquinoline

[0305] Compound 37b was prepared from 37a using the procedure describedfor 1b. ¹H—NMR 200 MHz (CDCl₃) δ: 7.57-7.88 (3H, m), 8.32 (1H, d, J=6Hz), 8.51-8.54 (1H, m).

[0306] 37c. 7-Bromo-1-phenoxyisoquinoline

[0307] Compound 37c was prepared from 37b using the procedure describedfor 1c. ¹H—NMR 200 MHz (CDCl₃) δ: 6.76-6.97 (2H, m), 7.18-8.09 (7H, m),8.60-8.64 (1H, m).

[0308] 37d. 1-Amino-7-bromoisoquinoline

[0309] Compound 37d was prepared from 37c using the procedure describedfor 1d. ¹H—NMR 200 MHz (CDCl₃) δ: 5.1 (2H, br. s), 7.03 (1H, dd, J=6 Hzand J=1 Hz), 7.59 (1H, d, J=9 Hz), 7.70 (1H, dd, J=9 Hz and J=2 Hz),7.95-8.00 (2H, m).

[0310] 37e. N-(7-bromo-1-isoquinolinyl)benzamide

[0311] Compound 37e was prepared from 37d using the procedure describedfor 1e. ¹H—NMR 200 MHz (CDCl₃) δ: 6.98 (1H,d, J=6 Hz), 7.37-7.57 (6H,m), 8.41-8.48 (2H, m), 9.14 (1H, d, J=2 Hz).

[0312] 37f. N-[7-(hydroxymethyl)-1-isoquinolinyl]benzamide

[0313] N-(7-Formylisoquinolinyl)benzamide was prepared from 37e usingthe procedure described for 1f but was not purified using columnchromatography. The crude aldehyde was transformed into the titlecompound using the procedure described for 1g folowed by purificationusing column chromatography on silica gel (toluene/ethyl acetate: 2/1).M.p. 137.5-139.0° C.

[0314] 37g.[[1-(benzoylamino)-7-isoquinolinyl]methyl][[(1,1-dimethylethoxy)carbonylamino]propane-dioicacid diethyl ester

[0315] Compound 37g was prepared from 37f using the procedure describedfor 1h.

[0316] M.p. 190.5-193.0° C.

[0317] 37h. 2-Amino-3-(1-amino-7-iaoquinolinyl)propionic aciddihydrochloride

[0318] Compound 37h was prepared from 37g using the procedure describedfor 1i. ¹H—NMR 200 MHz (D₂O) δ: 3.34-3.52 (2H, m), 4.29 (1H, dd, J=6 Hzand J=7 Hz), 7.13 (1H, dd, J=7 Hz and J=1Hz), 7.43 (1H, d, J=7 Hz), 7.82(2H, br.s), 8.05 (1H, br.s).

[0319] 37i. 2-Amino-3-(1-amino-7-isoquinolinyl)propionic acid methylester dihydrochloride

[0320] Compound 37i was prepared from 37h using the procedure describedfor 1j. ¹H—NMR 200 MHz (CD₃OD) δ: 3.38-3.60 (2H, m), 3.80 (3H, s), 4.52(1H, t, J=7 Hz), 7.25 (1H, dd, J=7 Hz and J=1 Hz), 7.59 (1H, d, J=7 Hz),7.87-7.99 (2H, m), 8.48 (1H, br.s).

[0321] 37j.1-[3-(1-amino-7-isoquinolinyl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]amino]-1-oxopropyl]-4-methylpiperidinehydrochloride

[0322] Using the procedure described for 11a, methyl ester 37i andcompound 12a gave3-(1-amino-7-isoquinolinyl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl)amino]propionicacid methyl ester. This ester was saponified and subsequently coupledwith 4-methylpiperidine using the procedure described for 4 to give 37.¹H—NMR 200 MHz (CD₃OD) δ: −0.20-0.77 (5H, m), 1.22-1.58 (3H, m),1.73-3.20 (4H, m), 3.65-4.19 (2H, m), 3.93 (3H, s), 4.53-4.68 (1H, m),7.00-7.06 (1H, m), 7.23-7.31 (2H, m), 7.43-7.51 (2H, m), 7.56-8.15 (6H,m).

Example 38(3S)-4-[[1-[(1-amino-7-isoquinolinyl)methyl]-2-oxo-2-(1-piperidinyl)ethyl]amino]-3-[[(4-methoxy-2,3,6-trimethylphenyl)sulfonyl]amino]-4-oxo-butanoicacid 1,1-dimethylethyl-ester hydrochloride

[0323] Using the procedure described for 5a aminoacid 37h gave1,1-dimethylethyl[1-[(1-amino-7-isoquinolinyl)methyl]-2-oxo-2-(1-piperidinyl)ethylcarbamate.This compound was deprotected and coupled with(2S)-2-[(4-methoxy-2,3,6-trimethylphenyl)sulfonyl]butanedioic acid4-(1,1-dimethylethyl)ester using the procedure described for example 8to give the title compound as a mixture of diastereoisomers (1.1).¹H—NMR 200 MHz (CD₃OD) δ: 1.30 and 1.33 (9H, 2× s), 1.38-1.68 (6H, m),2.12 and 2.14 (3H, 2× s), 2.25-2.43 (2H, m), 2.52 (3H, s), 2.61 (3H, s),2.90-3.56 (6H, m), 3.81 and 3.86 (3H, 2× s), 3.92-4.10 (1H, m),5.16-5.23 (1H, m), 6.72 and 6.75 (1H, 2× s), 7.18-7.24 (1H, m),7.51-7.56 (1H, m), 7.74-7.88 (2H, m), 8.23 and 8.26 (1H, 2× s).

Example 39 Solid-phase synthesis of compounds of formula (Id) with n=0,X=S, R¹Y=R¹C(O), R⁴=H (Table 39)

[0324] 39a. N-(4-thieno[3,2c]pyridinyl)acetylamide

[0325] 4.1 g of acetic anhydride was added to a solution of 5.0 g of 3bin 100 mL of pyridine at room temperature and the solution was heated at125° C. for 2.5 hours. The pyridine was evaporated, and the crudeproduct was coevaporated four times with toluene. The residue waschromatographed on a silica gel column (dichloromethane/methanol 95:5)to give 4.7 g of 39a.

[0326]¹H NMR 200 MHZ (CD₃OD) δ: 2.26 (3H,s), 7.49 (1H,d,J=6 Hz), 7.70(1H,d,J=6 Hz), 7.86 (1H,d,J=6 Hz), 8.21(1H,d,J=6 Hz).

[0327] 39b. N-(2-formylthieno[3,2c]pyridin-4-yl)acetylamide

[0328] The procedure described for 3d was used to prepare 39b from 39a.¹H NMR 200 MHZ (CD₃OD/CDCl₃9:1) δ: 2.33 (3H,s), 7.72 (1H,d), 8.30(1H,s), 8.32 (1H,d), 10.10 (1H,s).

[0329] 39c. N-[(4-(acetylamino)thieno[3,2c]pyridin-2-yl)methyl]-glycinemethyl ester

[0330] 1.7 g of 39b was dissolved in 80 mL of dioxane/methanol (1:1v/v). To this solution was added a solution of 1.0 g of glycine methylester hydrochloride and 1.1 g of N,N-diisopropylethylamine in methanol.The reaction mixture was heated at 60° C. for 1 hour, and thencoevaporated three times with methanol. Conversion of the aldehyde waschecked with NMR of the formed imine. ¹H NMR 200 MHZ (CD₃OD) δ: 2.30(3H,s), 3.79 (3H,s), 4.48 (2H,s), 7.75 (1H,d), 7.85 (1H,s), 8.25 (1H,d),8.60 (1H,s). The imine was dissolved in 40 mL of methanol and reduced tothe amine with sodium borohydride, added in small portions to an amountof 1.4 g. The reaction mixture was neutralized with acetic acid, thesolvents were evaporated, and the residue was coevaporated with toluene.The crude product was chromatographed on silica gel (toluene/ethanol9:1) giving 0.55 g of 39c. ¹H NMR 200 MHZ (CD₃OD) δ: 2.26 (3H,s), 3.47(2H,s), 3.70 (3H,s), 4.14 (2H,s), 7.32 (1H,s), 7.79 (1H,d), 8.19 (1H,d).

[0331] 39d.N,N-[tert.-butyloxycarbonyl][((4-(acetylamino)thieno[3,2c]pyridin-2-yl)methyl]-glycinemethyl ester.

[0332] The procedure described for 5a was used for the preparation of39d from 39c. ¹H NMR 200 MHZ (CD₃OD) δ: 1.47 (9H,s), 2.25 (3H,s), 3.69(3H,s), 4.02 (2H,m), 4.78 (2H,m), 7.37 (1H,m), 7.79 (1H,m), 8.20 (1H,m).

[0333] 39e.N,N-[tert-butyloxycarbonyl][(4-(acetylamino)thieno[3,2c]pyridin-2-yl)methyl]-glycine

[0334] Compound 39d was saponified as described for 2a but the reactionwas performed for 3 hours. Compound 39e was isolated by silica gelchromatography (dichloromethane/methanol 7:3). ¹H NMR 200 MHZ (CD₃OD) δ:1.48 (9H,s), 2.25 (3H,s), 3.84 (2H,m), 4.76 (2H,m), 7.33 (1H,m), 7.77(1H,m), 8.16 (1H,m).

[0335] 39f. Derivatization of Kaiser oxime resin with acid 39e.

[0336] 2.36 g of 39e was coevaporated twice with dryN,N-dimethylformamide and subsequently dissolved in 25 mL ofdichloromethane/N,N-dimethylformamide (3:2 v/v). 1.06 g ofN-hydroxybenzotriazole was added and the resulting solution was added to1.42 g of Kaiser oxime resin (1.1 mmol/g). After the addition of 1.22 mLof diisopropylcarbodiimide, the suspension was shaken overnight at roomtemperature. The resin was filtered off and washed withdichloromethane/N,N-dimethylformamide (3:2 v/v) andN,N-dimethylformamide. Further washings were performed by alternateaddition of 2-propanol and dichloromethane (three times each). Unreactedoxime functions were capped by treatment of the resin with 35 mL of amixture of aceticanhydride/N,N-diisopropylethylamine/N,N-dimethylformamide (3:1:12 v/v/v)for 30 minutes at room temperature. The resin was filtered off andwashed with N,N-dimethylformamide, 2-propanol and dichloromethane (threetimes each). The resin was dried in vacuo to give 1.9 g of 39f.

[0337] 39g.1-[2-[[(4-aminothieno[3,2c]pyridin-2-yl)methyl][benzoyl]amino]-1-oxoethyl]-4-methylpiperidine(compound of formula (Id) with n=0, X=S, R¹Y=benzoyl, R⁴=H,NR⁷R⁸=4-methylpiperidinyl)

[0338] 290 mg of 39f was treated with 6 mL of 25 vol % trifluoroaceticacid in dichloromethane for 30 minutes at room temperature. The resinwas filtered off and washed with dichloromethane, 2-propanol anddichloromethane. The resin was washed three times with 6 mLdichloromethane/N,N-dimethylformamide (3:2 v/v) containing 150 μlN,N-diisopropylethylamine and immediately reacted with 105 mg benzoicacid in 6 ml dichloromethane/N,N-dimethylformamide (3:2 v/v) containing150 μl of N,N-diisopropylethylamine and 400 mg ofbromotripyrrolidinophosphonium hexafluorophosphate (PyBrop). Thesuspension was shaken for 90 minutes at room temperature. The resin wasfiltered off and washed with dichloromethane/1methyl-2-pyrrolidinone(3:2 v/v), followed by washings with 1-methyl-2-pyrrolidinone,2-propanol and 1-methyl-2-pyrrolidinone. The reaction turned out to benot complete (chloranil test). The resin was reacted with 105 mg benzoicacid in 6 mL dichloromethane/N,N-dimethylformamide (3:2 v/v) containing150 μl of N,N-diisopropylethylamine and 400 mg of PyBrop. The suspensionwas shaken for 60 minutes at room temperature. The resin was filteredoff and washed with dichloromethane/1-methyl-2-pyrrolidinone (3:2 v/v),followed by washings with 1-methyl-2-pyrrolidinone, 2-propanol and1-methyl-2,-pyrrolidinone.The chloranil test revealed completeconversion.

[0339] 34 mg of the resin was suspended in 1 mL of a 0.5 M solution of4-methylpiperidine in distilled tetrahydrofuran and shaken for 16 hoursat room temperature. The resin was filtered off and washed withdichloromethane and methanol. The filtrates were collected andconcentrated to dryness. The residue was dissolved in 1 mLethylenediamine/ethanol (1:1 v/v) and shaken for 16 hours at roomtemperature. The reaction mixture was evaporated to dryness. The residuewas dissolved in dichloromethane, applied to a silica gel column, andeluted with a gradient dichloromethane/methanol=95/5v/v→dichloromethane/methanol=9/1 v/v. The UV positive fractions werepooled and evaporated to dryness yielding 7 mg of 39g.

Table 39

[0340] Solid-phase synthesis of compounds of formula (Id) with n=0, X=S,R¹Y=R¹C(O), R⁴=H

[0341] Using the procedure described for example 39g the carboxylicacids of structure R¹C(O)OH corresponding to R¹C(O) in Table 39 werecoupled to derivatized resin 39f. 34 mg portions of the resulting resinswere treated with amines of structure NHR⁷R⁸ as depicted in Table 39.When 3,4-dimethoxyaniline, 5-aminoindane or 4-aminobiphenyl was used asamine, the resin was suspended in 1 mL of a 0.5 M solution of amine indistilled tetrahydrofuran containing 2% acetic acid and was shaken for 3days. Work-up of the samples was performed as described for 39g.

[0342] All compounds were characterised by reversed phase liquidchromatography on a Supelcosil LC-18-DB column using followingconditions: Flow: 1.0 ml/min; Buffers A: water, B: acetonitrile/water(9:1 v/v), C: 0.5M phosphate buffer pH=2.1; Gradient : 0→45 min 75% A-5%B-20% C→15% A-65% B-20% C. UV-detection at 210 nm. Retention times aregiven in minutes in Table 39. TABLE 39 RP-HPLC retention times forexample 39

R¹C(O) R⁷R⁸N

23.2 25.8 28.6 32.9 18.4 24.6

25.5 27.6 30.2 34.7 20.9 26.5

15.6 18.0 21.1 24.4 11.3 16.7

29.5 31.3 33.5 38.4 25.2 30.2

31.0 32.7 35.1 39.4 27.3 31.9

26.2 28.2 30.9 34.6 22.2 27.3

35.0 35.7 38.5 43.0 34.7 35.5

38.6 39.4 41.6 45.7 35.6 39.0

Example 40(S)-[[1-(2-[[(1-amino-6-isoquinolinyl)carbonyl]amino]-1-oxopropyl)piperidin-4-yl]oxy]aceticacid hydrochloride

[0343] 40a. [1-(Benzoylamino)-6-isoquinolinyl]carboxylic acid

[0344] To a stirred solution of 400 mg sodium chlorite and 80 mg sodiumdihydrogen phosphate in 2.5 mL of water and 2.5 mL of dimethylsulfoxidein a cooling bath at room temperature was added dropwise a solution of510 mg N-(6-formyl-1-isoquinolinyl)benzamide (1f) in 2.5 mL ofdimethylsulfoxide. After 16 hours at room temperature a solution of 170mg sodium chlorite in 0.5 mL of water and 1.0 mL of dimethylsulfoxidewas added and stirred at room temperature for an additional 6 hours.Then 100 mL water was added, the pH adjusted to three using 2Nhydrochloric acid and the resulting suspension was kept at 5° C. for 16hours. The precipitate was collected to afford 450 mg of the titlecompound.

[0345] TLC: Rf=0.8, silica gel, ethyl acetate/pyridine/aceticacid/water: 63/20/6/11 v/v/v/v.

[0346] 40b. (1-Amino-6-isoquinolinyl)carboxylic acid hydrochloride

[0347] A mixture of 445 mg [1-(benzoylamino)-6-isoquinolinyl]carboxylicacid, 10 mL of acetic acid and 20 mL of 4N hydrochloric acid was heatedat 100° C. for one day. The reaction mixture was concentrated andcoevaporated with 0.5N hydrochloric acid. The resulting residue wastriturated with diethyl ether to yield 341 mg of the title compound

[0348]¹H—NMR 200 MHz (CD₃OD) δ: 7.35 (1H, dd, J=1 Hz and J=7 Hz), 7.63(1H, d, J=7 Hz), 8.30 (1H, dd, J=2 Hz and J=8 Hz), 8.49-8.57 (2H, m).

[0349] 40c. tert-Butyl(S)-[[1-(2-amino-1-oxopropyl)piperidin-4-yl]oxy]acetic acidhydrochloride

[0350] To a solution of 4.7 g of benzyl(S)-[2-[4-[(tert-butyloxycarbonyl)methoxy]piperidin-1-yl]-1-methyl-2-oxoethyl]carbamate(prepared from Z-L-Ala-OH as described in J. Med. Chem. 35, 4393 (1992)and EP0505868) in 80 mL of methanol were added 5 mL of 2N hydrochloricacid and 0.5 g of palladium on carbon (10%) and the mixture washydrogenated at atmospheric pressure. After two hours the mixture wasfiltered and the filtrate was concentrated to give 3.3 g of the titlecompound.

[0351] TLC: Rf=0.3, silica gel, ethyl acetate/pyridine/aceticacid/water: 63/20/6/11 v/v/v/v.

[0352] 40d. tert-Butyl(S)-[[1-(2-[[(1-amino-6-isoquinolinyl)carbonyl]amino]-1-oxopropyl)piperidin-4-yl]oxy]aceticacid

[0353] To a solution of 0.33 g (1-amino-6-isoquinolinyl)carboxylic acidhydrochloride in 20 mL of N,N-dimethylformamide were added 0.34 ghydroxybenztriazole, 0.504 mL N-methylmorpholine, 0.525 g tert-butyl(S)-[[1-(2-amino-1-oxopropyl)piperidin-4-yl]oxy]acetic acidhydrochloride and 0.425 g 1-(3-dimethylaminopropyl)-3-ethylcarbodiimideAfter stirring at room temperature for three days the mixture wasconcentrated. Dichloromethane and 1% aqueous sodium hydrogencarbonatewere added to the residue and the organic layer was separated. Theaqueous layer was washed four times with dichloromethane, the combinedorganic layers dried (sodium sulfate) and concentrated. The residue waspurified by chromatography (silica gel, dichloromethane/methanol: 10/1v/v followed by a purification using silica gel, ethyl acetate/methanol:10/1 v/v) to give 0.619 g of the title compound.

[0354] TLC: Rf=0.4, silica gel, dichloromethane/methanol: 10/1 v/v.

[0355] 40e.(S)-[[1-(2-[[(1-amino-6-isoquinolinyl)carbonyl]amino]-1-oxopropyl)piperidin-4-yl]oxy]aceticacid hydrochloride

[0356] To a solution of 594 mg of tert-butyl(S)-[[1-(2-[[(1-amino-6-isoquinolinyl)carbonyl]amino]-1-oxopropyl)piperidin-4-yl]oxy]aceticacid in 8 mL of dioxane was added 3 mL 36% hydrochloric acid and stirredat room temperature for two hours. The solution was concentrated andtrituration of the residue with diethyl ether yielded 560 mg of thetitle compound. HPLC Supelcosil LC-18-DB column using a gradient elutionsystem of 20% A/80% B to 20% A/20% B/60% C over 40 min at a flow of 0.25ml/min(A: 0.5M phosphate buffer pH 2.1, B: water, C acetonitril/water3/2 v/v). Rt=22.4 min.

Example 41 Ethyl(S)-[[1-(2-[[1(1-amino-6-isoquinolinyl)carbonyl]amino]-1-oxopropyl)piperidin-4-yl]oxy]aceticacid hydrochloride

[0357] To a stirred solution of 394 mg of(S)-[[1-(2-[[(1-amino-6-isoquinolinyl)carbonyl]amino]-1-oxopropyl)piperidin-4-yl]oxy]aceticacid hydrochloride in 10 mL of ethanol at 0° C. was added 0.55 mL ofsulfuric acid (95-98%). The reaction mixture was allowed to warm to roomtemperature and after 2 hours 5 mL of 2N aqueous sodium hydroxide, 20 mLof brine, 20 mL of 5% aqueous sodium hydrogencarbonate and 40 mL ofdichloromethane was added. The organic layer was separated and theaqueous layer was extracted three times with dichloromethane. Thecombined organic layers were dried (sodium sulfate) and concentrated.Purification using column chromatography (silica gel, dichloromethane:ethanol=9:1) and lyophilisation (t-butanol/hydrochloric acid) yielded265 mg of the title compound.

[0358] Rt=27.1 min on HPLC Supelcosil LC-18-DB column using a gradientelution system of 20% A /80% B to 20% A/20% B/60% C over 40 min at aflow of 0.25 ml/min (A: 0.5M phosphate buffer pH 2.1, B: water, Cacetonitril/water 3/2 v/v).

Example 421-[3-(4-Aminofuro[3,2c]pyridin-2-yl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]amino]-1-oxopropyl]-4-methylpiperidinehydrochloride (42h)

[0359] 42a. 4-Aminofuro[3,2c]pyridine

[0360] Liquid ammonia (150 mL) was added to a solution of 15.3 g (100mmol) 4-chlorofuro[3,2c]pyridine (J. S. New et al., J.Med. Chem. 32,1147 (1989)) in 550 mL of ethanol in a steel vessel. Nitrogen waspressed upon until an initial pressure of 4 atm was obtained. Thisreaction mixture was heated for 2 days at 200° C. The solvent wasremoved in vacuo and the residue dissolved in water The pH value wasadjusted to pH 10 by adding aqueous sodium carbonate solution, followedby extraction with ethyl acetate. The organic extract was washed withbrine and dried (sodium sulfate). Evaporation of the solvent in vacuogave pure 4-aminofuro[3,2c]pyridine. Yield: 12.2 g (91%); m.p. 120-122°C.; EI-MS. 134 (M⁺).

[0361] 42b. N-(furo[3,2c]pyridin-4-yl)benzamide

[0362] The procedure described for 1e was used to prepare 42c from 42b.¹H—NMR 200 MHz (CDCl₃) δ: 7.20-7.69 (6H, m), 8.17-8.87 (3H, m).

[0363] 42c. N-[2-(hydroxymethyl)furo[3,2c]pyridin-4-yl]benzamide

[0364] To a stirred solution of 1.27 mL of n-butyl lithium (1.6 M inhexane) in 4 mL of tetrahydrofuran under a nitrogen atmosphere at −78°C. was added dropwise a solution of 81 mg ofN-(4-furo[3,2c]pyridinyl)benzamide in 10 mL of tetrahydrofuran over aperiod of 15 minutes. After stirring for 20 min a mixture of 0.5 mL ofN,N-dimethylformamide and 2 mL of tetrahydrofuran was added fast. Thecooling bath was removed, the reaction mixture was allowed to come to 0°C. and poured into a cold mixture of 1 mL of 2 N hydrochloric acid and50 mL of brine. The mixture was adjusted to pH 6 and extracted withethyl acetate. The ethyl acetate extract was dried (magnesium sulfate)and concentrated under reduced pressure. The residue was dissolved in 1mL of tetrahydrofuran and 4 mL of methanol and 10 mg of sodiumborohydride was added in small portions. After stirring the mixture atambient temperature for 10 min, 10 mL of water was added, the pHadjusted to 8 using 1N hydrochloric acid and tetrahydrofuran andmethanol were removed in vacuo. Brine was added and the mixture wasextracted with ethyl acetate. The ethyl acetate extract was dried(magnesium sulfate) and concentrated under reduced pressure. The crudeproduct chromatographed on a silica gel column (toluene/ethyl acetate:1/2) yielding 45 mg of title compound 42c. ¹H—NMR 200 MHz (CDCl₃) δ:4.74 (2H, s), 6.92-6.95 (1H, m), 7.30-7.68 (4H, m), 7.99-8.18 (3H, m).

[0365]42d.[[4-(benzoylamino)furor[3,2c]pyridin-2-yl]methyl][[(1,1-dimethylethoxy)carbonyl]amino]-propanedioicacid diethyl ester

[0366] The procedure described for 1h was used to prepare 42d from 42c.Purification using column chromatography on silica gel (toluene: ethylacetate=4:1) afforded 16% of title compound 42d. ¹H—NMR 200 MHz (CDCl₃)δ: 1.31 (6H, t, J=7 Hz), 1.46 (9H, s), 3.89 (2H, s), 4.25-4.38 (4H, m),6.90 (1H, br.s), 7.10-7.63 (4H, m), 7.96-8.10 (3H, m),

[0367] Elution of the column with ethyl acetate yielded a mixture whichwas rechromatographed on silica gel (ethyl acetate) giving 6% of[(4-aminothieno[3,2c]pyridin-2-yl)methyl][[(1,1-dimethylethoxy)carbonyl]amino]propanedioicacid diethyl ester. ¹H—NMR 200 MHz (CDCl₃) δ: 1.30 (6H, t, J=7 Hz), 1.46(9H, s), 3.85 (2H, s), 4.23-4.38 (4H, m), 6.35 (1H, d, J=1 Hz), 6.75(1H, dd, J=1 Hz and J=6 Hz), 7.87 (1H, d, J=6 Hz).

[0368] 42e.2-Amino-3-(4-aminofuro[3,2c]pyridin-2-yl)propionic aciddihydrochloride

[0369] This compound was prepared from[2-[4-(benzoylamino)furo[3,2c]pyridinyl]methyl][[(1,1-dimethylethoxy)carbonyl]amino]propanedioic acid diethyl ester and[(4-aminothieno[3,2c]pyridin-2-yl)-methyl][[(1,1-dimethylethoxy)carbonyl]amino]propanedioicacid diethyl ester using the procedure described for 1i. NMR 200 MHz(D₂O) δ: 3.55 (2H, d, J=6 Hz), 4.44 (1H, t, J=6 Hz), 7.00 (1H, d, J=1Hz), 7.11 (1H, dd, J=1 Hz and J=7 Hz) 7.66 (1H,d, J=7 Hz).

[0370] 42f.2-Amino-3-(4-aminofuro[3,2c]pyridin-2-yl)propionic acidmethyl ester dihydrochloride

[0371] This compound was prepared from 42e using the procedure describedfor 1j. ¹H—NMR 200 MHz (CD₃OD) δ: 3.59 (2H, d, J=6 Hz), 3.90 (3H, s),4.57 (1H, t, J=6 Hz), 7.21 (1H, d, J=7 Hz), 7.22 (1H, s), 7.79 (1H, d,J=7 Hz).

[0372]42g.3-(4-Aminofuro[3,2c]pyridin-2-yl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]amino]propionicacid methyl ester

[0373] Using the procedure described for 11a, the reaction of methylester 42f and sulfonyl chloride 12a gave the tile compound ¹H—NMR 200MHz (CDCl₃ and 20% MeOD) δ: 3.02-3.26 (2H, m), 3.49 (3H, s), 3.94 (3H,s), 4.35 (1H, dd, J=5 Hz and J=9 Hz), 6.46-6.51 (2H, m), 7.16 (1H, d,J=2.5 Hz), 7.25 (1H, dd, J=2.5 Hz and J=9 Hz), 7.45-7.74 (4H, m), 8.17(1H, d, J=2 Hz).

[0374]42h.1-[3-(4-Aminofuro[3,2c]pyridin-2-yl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]amino]-1-oxopropyl]-4-methylpiperidinehydrochloride

[0375] Compound 42g was saponified and subsequently coupled with4-methylpiperidine using the procedure described for 4 to give 42h.¹H—NMR 400 MHz (CD₃OD) δ: 0.30-0.41 (0.6H, m), 0.61-0.84 (1.4H, m), 0.66( 1.8H, d, J=6 Hz), 0.81 (1.2H, d, J=1.2H), 1.37-1.68 (3H, m), 2.15-2.23(0.4H, m), 2.41-2.50 (0.6H, m), 2.88-3.14 (3H, m), 3.86-3.95 (1H, m),3.96 (3H, s), 4.07-4.22 (1H, m), 4.68-4.78 (1H, m), 6.80 (0.4H, s), 6.85(0.6H, s), 6.93 (0.4H, d, J=7 Hz), 6.97 (0.6H, d, J=7 Hz), 7.27-7.31(2H, m), 7.49-7.58 (2H, m), 7.77-7.82 (2H, m), 8.13 (0.4H, d, J=2 Hz),8.15 (1H, d, J=2 Hz).

Example 431-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(2-trifluoroacetyl-1,2,3,4-tetrahydro-7-isoquinolinyl)sulfonyl]amino]-1-oxopropyl]-4-methylpiperidine (43c)

[0376] 43a. 2-Trifluoroacetyl-1,2,3,4-tetrahydroisoquinoline

[0377] This intermediate was prepared using the experimental proceduredescribed for 3 trifluoroacetyl-2,3,4,5-tetrahydro-1H-3-benzazepine(preparation 3b) in WO 95/13274). The crude product was distilled atreduced pressure (p=1.2 mbar, T=95-110° C.) yielding 77% of titlecompound 43a.

[0378] 43b.(2-trifluoroacetyl-1,2,3,4-tetrahydro-7-isoquinolinyl)sulfonyl chloride

[0379] This compound was obtained by the method described in J.Med.Chem.23, 837 (1980). Crystallization from ether yielded 65% of title compound43b (rf=0.34; silica gel: heptane/ether=4/6). 43c.1-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(2-trifluoroacetyl-1,2,3,4-tetrahydro-7-isoquinolinyl)sulfonyl]amino]-1-oxopropyl]-4-methylpiperidine

[0380] To 0.56 g of 1,1-dimethylethyl[1-[(4-aminothieno[3,2c]pyridin-2-yl)methyl]-2-oxo-2-[1(4-methylpiperidinyl)ethylcarbamate(30a) in 16 mL of dichloromethane was added 16 mL of trifluoroaceticacid. After stirring for 1.5 hours at room temperature the reactionmixture was concentrated and coevaporated with toluene. The residue wasdissolved in 20 mL of dichloromethane and 0.56 mL of triethylamine,cooled at 0° C. and 0.49 g of(2-trifluoroacetyl-1,2,3,4-tetrahydro-7-isoquinolinyl)sulfonyl chloridein 8 mL of dichloromethane was added dropwise. The pH of the reactionmixture was adjusted to 8-9 with triethylamine (an additional 0.22 mLwas required). After stirring for 0.5 hours at room temperature waterwas added, the pH was adjusted to 8-9 and the mixture was extracted withdichloromethane. The extract was dried (magnesium sulfate) andconcentrated. Purification on silica gel (dichloromethane/methanol: 95/5v/v) yielded 0.69 g of title compound 43c. ¹H—NMR 200 MHz (CDCl₃) δ:0.62-0.93 (4H, m), 1.35-1.68 (3H, m) 2.18-3.30 (11H, m), 3.60-3.92 (3H,m), 4.18-4.62 (2H, m), 4.70-4.77 (2H, m), 6.98-7.30 (3H, m), 7.52-7.62(2H, m), 7.72-7.80 (1H, m).

Example 441-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(1,2,3,4-tetrahydro-7-isoquinolinyl)sulfonyl-amino]-1-oxopropyl]-4-methylpiperidine

[0381] Using the experimental procedure described in “preparation 23” ofpatent WO 95/13274 (Pfizer) 0.64 g of 43c yielded 0.43 g of 44 aftercrystallization from ethanol. ¹H—NMR 200 MHz (CDCl₃, 10% CD₃OD) δ:0.05-0.98 (4H, m), 1.34-1.68 (3H, m), 2.15-2.58 (1H, m), 2.64-3.77 (12H,m), 3.91-3.99 (2H, m), 4.19-4.38 (1H, m), 4.45-4.57 (1H, m), 7.01-7.17(3H, m), 7.39-7.53 (2H, m), 7.74 and 7.78 (1H, 2× s).

Example 451-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[[2-(2-propyl)-1,2,3,4-tetrahydro-7-isoquinolinyl]sulfonyl]amino]-1-oxopropyl]-4-methylpiperidinedihydrochloride

[0382] To 91 mg of 44 dissolved in 5 mL of tetrahydrofuran under anitrogen atmosphere was added 127 mg of potassium carbonate and 90 μl of2-iodopropane. After stirring at 65° C. for one day an additional 120 mgof potassium carbonate and 90 μl of 2-iodopropane were added and thereaction mixture stirred at 65° C. for an additional day. The solventwas evaporated and dichloromethane and water were added. The organiclayer was separated, dried (magnesium sulfate) and concentrated.Purification by chromatography on silica gel using dichloromethane:methanol=85:15 (v/v) yielded 41 mg of free base. Treatment of this freebase with 2 equivalents hydrogen chloride and lyophilisation yielded thetitle compound. ¹H—NMR 400 MHz (CD₃OD) δ: 0.68 and 0.94 (3H, 2× t, J=6Hz), 1.46 (6H, d, J=7 Hz), 4.62 (1H, t, J=7 Hz), 7.30-7.39 (2H, m),7.54-7.72 (4H, m), 0.21-4.52 (remaining protons, m).

Example 461-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(2-methylsulfonyl-1,2,3,4-tetrahydro-7-isoquinolinyl)sulfonyl]amino]-1-oxopropyl]-4-methylpiperidinehydrochloride

[0383] A mixture of 99 mg of 44, 4 mL of dichloromethane and 0.98 mL oftriethylamine was cooled at 0° C. A total of 0.43 mL ofmethanesulfonylchloride was added in small quantities and the reactionmixture was stirred 8 hours at 0° C. The reaction mixture was dilutedwith dichloromethane and washed with water, dried (magnesium sulfate)and concentrated. Purification by chromatography on silica gel usingdichloromethane: methanol=9:1 yielded free base. Treatment of this freebase with one equivalent hydrogen chloride and lyophilisation yielded 88mg of title compound 46. ¹H—NMR 400 MHz (CD₃OD) δ: 0.30-0.94(2H, m),0.71 and 0.91 (3H, 2× t, J=6 Hz), 1.45-1.66 (3H, m), 2.23-3.20 (6H, m),2.91 and 2.93 (3H, 2× s), 3.47-3.58 (2H, m), 3.75-3.93 (1H, m),4.11-4.32 (1H, m), 4.38 and 4.42 (2H, 2× s), 7.24-7.40 (2H, m),7.51-7.61 (4H, m)

Example 471-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(2-trifluoroacetyl-1,2,3,4-tetrahydro-6-isoquinolinyl)sulfonyl]amino]-1-oxopropyl]-4-methylpiperidinehydrochloride (47b)

[0384] 47a.(2-trifluoroacetyl-1,2,3,4-tetrahydro-6-isoquinolinyl)sulfonyl chloride

[0385] The mother liquor obtained in the synthesis of 43b was subjectedto column chromatography on silica gel (heptane/ether=4/6) affordingtitle compound 47a (rf=0.46; SiO₂: heptane/ether=4/6).

[0386] 47b.1-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(2-trifluoroacetyl-1,2,3,4-tetrahydro-6-isoquinolinyl)sulfonyl]amino]-1-oxopropyl]-4-methylpiperidinehydrochloride

[0387] Using the experimental procedure described for 43c, 30a wasdeprotected and coupled with 47a to yield compound 47b. ¹⁹F—NMR 188 MHz(CD₃OD) δ: −71.7.

Example 481-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(4-methoxy-2,3,6-trimethylphenyl)sulfonyl]-amino]-1-oxopropyl]-4-methylpiperidinehydrochloride

[0388] The procedure described for 30b was used to couple 30a with4-methoxy-2,3,6-trimethylphenylsulfonylchloride to give the titlecompound. ¹H—NMR 400 MHz (CD₃OD) δ: 0.38-0.97 (2H, m), 0.74 and 0.88(3H, 2× t, J=7 Hz), 1.29-1.62 (3H, m), 1.99 and 2.01 (3H, 2× s),2.27-3.3 (4H, m), 2.46 (3H, s), 2.50 (3H, s),3.65-3.81 (1H, m), 3.81 and3.82 (3H, 2× s), 4.19-4.47 (2H, m), 6.61 (1H, s), 7.31-7.34 (1H, m),7.48-7.50 (1H, m), 7.56-7.60 (1H, m)

Example 494-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]amino]-1-oxopropyl]-N,N-dimethylamino-1-piperazinecarboxamidehydrochloride (49b)

[0389] 49a. N,N-dimethylamino-1-piperazinecarboxamide hydrochloride.

[0390] To a solution of 1.92 g1-(1,1-dimethylethyloxycarbonyl)piperazine in 11 mL of dichloromethaneand 1.5 mL of triethylamine at room temperature w,as added dropwise 1 mLof dimethylcarbamylchloride. After 16 hours 5% aqueous sodiumhydrogencarbonate and dichloromethane were added, the organic layerseparated, dried over magnesium sulfate and concentrated. Columnchromatography on silica gel (dichloromethane/methanol=95/5 v/v) yielded1.34 g1-(1,1-dimethylethyloxycarbonyl)-4-(dimethylaminocarbonyl)piperazine.This compound was dissolved in a 3N hydrogen chloride solution inmethanol at 0° C. After 15 min the reaction mixture was allowed to warmto room temperature and After stirring for 3 days at room temperaturethe reaction mixture was concentrated to give 1.06 g of1-(dimethylaminocarbonyl)piperazine hydrochloride. rf=0.29, SiO₂:dichloromethane/methanol=4/1 v/v.

[0391] 49b.4-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(7-methoxy-2-naphthalenyl)sulfonyl]amino]-1-oxopropyl]-N,N-dimethylamino-1-piperazinecarboxamidehydrochloride

[0392] Using the procedure described for 2b, 49a was coupled with 14a togive compound 49b. ¹H—NMR 200 MHz (CD₃OD) δ: 2.55-3.64 (6H, m), 2.78(6H, s), 3.94 (3H, s), 4.63 (1H, dd, J=9 Hz and J=5 Hz), 7.20-7.59 (6H,m), 7.76-7.84 (2H, m), 8.15-8.18 (1H, m).

Example 501-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(7-nitrodibenzofuran-2-yl)sulfonyl]amino]-1-oxopropyl]-4-methylpiperidinehydrochloride (50b)

[0393] 50a. (7-Nitrodibenzofuran-2-yl)sulfonylchloride

[0394] To a stirred solution of 100 mg of 3-nitrodibenzofuran in 1.7 mLof dichloromethane at −20° C. under a nitrogen atmosphere was added 0.44mL chlorosulfonic acid in small portions. The reaction mixture wasallowed to slowly warm to room temperature and stirred for 66 hours atroom temperature. The reaction mixture was poured into ice-cold water,extracted four times with dichloromethane, dried (magnesium sulfate) andconcentrated. Purification on silica gel (dichloromethane) yielded 129mg of 50a: rf=0.73.

[0395] 50b.1-[3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(7-nitrodibenzofuran-2-yl)sulfonyl]amino]-1-oxopropyl]-4-methylpiperidinehydrochloride

[0396] The procedure described for 30b was used to couple 30a with 50agiving title compound 50b.

[0397]¹H—NMR 400 MHz (CDCl₃) δ: 0.39 and 0.81 (3H, 2× d, J=6 Hz),0.06-3.34 (9H, m), 3.88-4.19 (m, 2H), 4.71-4.79 (1H, m), 6.82-8.86 (12H,m).

Example 51(2S)-N-[1-[(1-amino-6-isoquinolinyl)methyl]-2-oxo-2-(1-piperidinyl)ethyl]-4-(4-morpholinyl)-4-oxo-2-[(4-methoxy-2,3,6-trimethylphenylsulfonyl)amino]butanamidehydrochloride

[0398] 51a.(2S)-4-(4-Morpholinyl)-4-oxo-2-[(4-methoxy-2,3,6-trimethylphenylsulfonyl)amino]butanoicacid

[0399] 2.45 g of Fmoc-Asp-OtBu was dissolved in 10 mL of dichloromethaneand 0.63 mL of morpholine and 2.05 g of2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroboratewere added. The pH of the reaction mixture was kept at 8 usingtriethylamine. After stirring 1 hour at room temperature aqueous 5%sodium hydrogencarbonate was added to the reaction mixture. The organiclayer was separated, washed with water, dried over magnesium sulfate andconcentrated. The residue was dissolved in 20 mL ofN,N-dimethylformamide and 5 mL of piperidine was added. After stirring 1hour at room temperature the reaction mixture was concentrated, theresidue was dissolved in ethyl acetate and extracted several times withice-cold water adjusted to pH 3 with 1N hydrochloric acid. The combinedwater layers were saturated with sodium chloride, made basic (pH 9)using 2N sodium hydroxide and extracted with dichloromethane. Thecombined dichloromethane layers were dried over magnesium sulfate andconcentrated. The residue was dissolved in 30 mL of dichloromethane and1.4 mL of triethylamine and 2.5 g of4-methoxy-2,3,6-trimethylphenylsulfonylchloride (Mtr-chloride) wereadded. After stirring 2 hours at room temperature aqueous 5% sodiumhydrogencarbonate was added to the reaction mixture and extracted threetimes with dichloromethane. The combined dichloromethane layers weredried over magnesiumsulfate and concentrated. The residue was dissolvedin 40 mL of dichloromethane, 10 mL of trifluoroacetic acid was added andstirred at room temperature for one hour. The reaction mixture wasconcentrated and coevaporated twice with toluene. Dichloromethane andwater were added to the residue and the mixture was made basic (pH 9)using aqueous 2N sodium hydroxide. The aqueous layer was separated andwashed with dichloromethane. The dichloromethane layers were washed withaqueous 5% sodium hydrogencarbonate. The combined basic aqueous layerswere made acid (pH 2) using 2 N hydrochloric acid and three timesextracted with dichloromethane. The combined dichloromethane layers weredried over magnesium sulfate and concentrated to give 1.94 g of compound51a. ¹H—NMR 200 MHz (CDCl₃) δ: 2.14 (3H, s), 2.60 (3H, s), 2.65 (3H, s),2.83-3.98 (11H, m), 3.85 (3H, s), 6.05 (1H, d), 6.57 (1H, s).

[0400]51b.(2S)-N-[1-[(1-amino-6-isoquinolinyl)methyl]-2-oxo-2-(1-piperidinyl)ethyl]-4-(4-morpholinyl)-4-oxo-2-[(4-methoxy-2,3,6-trimethylphenylsulfonyl)amino]butanamidehydrochloride

[0401] The procedure described for 5c was used. Deprotection of 5a andcoupling with 51a yielded after purification the title compound (79%) asa mixture of diastereomers (1:1). ¹H—NMR 200 MHz (CD₃OD) δ: 1.30-1.51(6H, m), 2.14 (3H, s), 2.51 and 2.53 (3H, 2× s), 2.63 (3H, s), 2.31-3.62(16H, m), 3.84 and 3.86 (3H, 2× s), 3.99-4.15 (1H, m), 5.08-5.21 (1H,m), 6.75 and 6.76 (1H, 2× s), 7.18-7.25 (1H, m), 7.52-7.81 (3H, m),8.25-8.37 (1H, m).

Example 52(4S)-5-[[1-[(1-amino-6-isoquinolinyl)methyl]-2-oxo-2-(1-piperidinyl)ethyl]amino]-4-[[(4-methoxy-2,3,6-trimethylphenyl)sulfonyl]amino]-5-oxo-pentanoicacid ethylester hydrochloride

[0402] The procedure described for 5c was used. Deprotection of 5a andcoupling with Mtr-Glu(OEt)-OH (prepared from Fmoc-Glu-OtBu, ethanol andMtr-chloride according to the procedure described for 51a) yielded afterpurification the title compound (76%) as a mixture of diastereomers(1:1). ¹H—NMR 400 Hz (CD₃OD) δ: 1.15-2.32 (11H, m), 2.13 (3H, s), 2.40and 2.42 (3H, 2× s), 2.44 and 2.47 (3H, 2× s), 2.75-4.08 (11H, m),4.99-5.19 (1H, m), 6.71 and 6.72 (1H, 2× s), 7.16-7.20 (1H, m),7.51-7.76 (3H, m), 8.30-8.36 (1H, m).

Example 53(3S)-4-[[4-[[1-[[(1-amino-6-isoquinolinyl)methyl]-2-oxo-2-(1-piperidinyl]amino]-3-[[(4-methoxy-2,3,6-trimethylphenyl)sulfonyl]amino]-1,4-dioxobutyl]amino]butanoicacid ethyl ester hydrochloride

[0403] The procedure described for 5c was used. Deprotection of 150 mgof 5a and coupling with 189 mg of Mtr-Asp(NH—(CH₂)₃COOEt)-OH (preparedfrom Fmoc-Asp-OtBu and 4-aminobutanoic acid ethyl ester using theprocedure described for 51a) yielded after purification compound 53 (155mg) as a mixture of diastereomers (1:1). ¹H—NMR 400 MHz (CD₃OD) δ: 1.22(3H, t, J=7 Hz), 1.05-1.74 (8H, m), 2.12 (3H, s), 2.24-2.48 (4H, m),2.53 (3H, s), 2.62 (3H, s), 2.89-3.56 (8H, m), 3.81 (3H, s), 3.85 (3H,s), 3.99-4.13 (3H, m), 5.02-5.16 (1H, m), 6.72 and 6.73 (1H, 2× s),7.11-7.15 (1H, m), 7.53-7.70 (3H, m), 8.21-8.26 (1H, m).

Example 54(2S)-N-[1-[(1-amino-6-isoquinolinyl)methyl]-2-oxo-2-(1-piperidinyl)ethyl]-2-[[(4-methoxy-2,3,6-trimethylphenyl)sulfonyl]amino]-4-methylpentanamidehydrochloride

[0404] The procedure described for 5c was used. Deprotection of 75 mg of5a and coupling with 72 mg of(2S)-2-[[(4-methoxy-2,3,6-trimethylphenyl)sulfonyl]amino]-4-methylpentanoicacid (prepared from L-leucine and4-methoxy-2,3,6-trimethylphenylsulfonylchloride using the proceduredescribed for 5b) yielded after purification the title compound (65 mg)as a mixture of diastereomers (1:1). ¹H—NMR 400 MHz (CD₃OD) δ: 0.50 and0.68 (3H, 2× d, J=7 Hz), 0.61 and 0.79 (3H, 2× d, J=7 Hz), 0.97-1.65(9H, m), 2.13 (3H, s), 2.54 and 2.56 (3H, 2× s), 2.61 and 2.62 (3H, 2×s), 2.76-3.68 (7H, m), 3.78 and 3.84 (3H, 2× s), 4.98-5.02 and 5.18-5.22(1H, 2× m), 6.71 (1H, s), 7.16-7.19 (1H, m), 7.53-7.78 (4H, m),8.29-8.34 (1H, m).

Example 55(2S)-N-[1-[(1-amino-6-isoquinolinyl)methyl]-2-oxo-2-(1-piperidinyl)ethyl]-3-phenyl-2-[[(4-methoxy-2,3,6-trimethylphenyl)sulfonyl]amino]propanamidehydrochloride

[0405] To a solution of 0.5 g of L-phenylalanine t-butyl esterhydrochloride in 4 mL of N,N-dimethylformamide were added 0.67 g of4-methoxy-2,3,6-trimethylphenylsulfonylchloride and 0.96 mL ofN,N-diisopropylethylamine. After stirring for 2 hours at roomtemperature the reaction mixture was concentrated and the residuedissolved in ethyl acetate. The ethyl acetate solution was washed withaqueous potassium hydrogensulfate (5%), water, aqueous sodiumhydrogencarbonate (5%) and brine, dried over magnesium sulfate andconcentrated. The residue was dissolved in 16 mL of dichloromethane and4 mL of trifluoroacetic acid was added. After stirring for 2 h at roomtemperature the reaction mixture was concentrated. Dichloromethane andaqueous sodium hydrogencarbonate (5%) were added to the residue (themixture was basic), the aqueous layer separated and washed withdichloromethane. The aqueous layer was made acid (pH 2) using 2 Nhydrochloric acid and several times extracted with dichloromethane. Thecombined dichloromethane layers were washed with brine, dried overmagnesium sulfate and concentrated to give 0.65 g of(2S)-2-[[(4-methoxy-2,3,6-trimethylphenyl)sulfonyl]amino]-3-phenylpropanoicacid. This acid (80 mg) was coupled with deprotected 5a (75 mg)according to the procedure described for 5c to afford compound 55 (55mg) as a mixture of diastereomers (1:1). ¹H—NMR 400 MHz (CD₃OD) δ:1.25-1.67 (6H, m), 1.97 and 1.99 (3H, 2× s), 2.12 and 2.17 (3H, 2× s),2.45-3.54 (8H, m), 2.49 (3H, s), 3.81 and 3.85 (3H, 2× s), 3.82-3.92(1H, m), 5.13-5.28 (1H, m), 6.61 (1H, s), 6.84-7.07 (5H, m), 7.17-7.21(1H, m), 7.50-7.56 (1H, m), 7.65-7.69 (1H, m), 7.76-7.82 (1H, m),8.30-8.35 (1H, m).

Example 56(3S)-4-[[1-[(1-amino-6-isoquinolinyl)methyl]-2-oxo-2-(1-piperidinyl)ethyl]amino]-3-[[(phenylmethyl)sulfonyl]amino]-4-oxo-butanoicacid 1,1-dimethylethylester hydrochloride

[0406] The procedure described for 5c was used. Deprotection of 100 mgof 5a and coupling with 95 mg of(²S)-[[(phenylmethyl)sulfonyl]amino]butanedioic acid4-(1,1-dimethylethyl)ester (prepared from Asp(OtBu)-OH andphenylmethylsulfonylchloride using the procedure described for 5b)yielded the title compound (133 mg) as a mixture of diastereomers (1:1).¹H—NMR 400 MHz (CD₃OD) δ: 0.83-1.69 (6H, m), 1.41 and 1.42 (9H, 2× s),2.42-2.63 (2H, m), 3.09-3.61 (6H, m), 4.01-4.38 (3H, m), 5.22-5.31 (1H,m), 7.13-7.80 (9H, m), 8.25-8.30 (1H, m).

Example 57(3S)-4-[[1-[(1-amino-6-isoquinolinyl)methyl]-2-oxo-2-(1-piperidinyl)ethyl]amino]-3-[[4-methoxyphenyl)sulfonyl]amino]-4-oxo-butanoicacid 1,1-dimethylethylester hydrochloride

[0407] The procedure described for 5c was used. Deprotection of 100 mgof 5a and coupling with 99 mg of(2S)-[[(4-methoxyphenyl)sulfonyl]amino]butanedioic acid4-(1,1-dimethylethyl)ester (prepared from Asp(OtBu)-OH and4-methoxyphenylsulfonylchloride using the procedure described for 5b)yielded the title compound (85 mg) as a mixture of diastereomers (1:1).¹H—NMR 400 MHz (CD₃OD) δ: 1.25-1.67 (6H, m), 1.32 and 1.35 (9H, 2× s),2.18-2.48 (2H, m), 3.96-3.55 (6H, m), 3.83 and 3.86 (3H, 2× s),4.04-4.14 (1H, m), 5.05-5.17 (1H, m), 6.99-7.05 (2H, m), 7.19-7.23 (1H,m), 7.51-7.79 (5H, m), 8.30-8.36 (1H, m).

Example 58N-[1-(4-Amino-thieno[3,2-c]pyridin-2-ylmethyl)-2-(4-methyl-piperidin-1-yl-2-oxo-ethyl]-4-(tetrahydropyran-4-yloxy)-benzenesulfonamide(58c)

[0408] 58a. 4-(4-Bromo-phenoxy)-tetrahydropyran

[0409] A mixture of 4.0 mL of diethyl-azodicarboxylate in 20 mL of drytetrahydrofuran was added at 5° C. to a stirred solution of 3.5 g of4-bromo-phenol, 2.4 mL of 4-hydroxy-tetrahydropyran and 6.6 gtriphenylphosphine in 75 mL of tetrahydrofuran within 30 minutes.Stirring was continued at room temperature for 72 hours. The solvent wasevaporated in vacuo and the residue chromatographed on silica gel (ethylacetate) yielding 5.6 g of 4-(4-bromo-phenoxy)-tetrahydropyran as awhite solid. M.p 53-55° C., EI-MS: 256 (M⁺).

[0410] 58b. 4-(Tetrahydropyran-4-yloxy)-benzenesulfonyl chloride

[0411] To a solution of 2.8 g of 4-(4-Bromo-phenoxy)-tetrahydropyran in75 mL of dry tetrahydrofuran was added 7.5 mL of n-butyllithium (1.6 Nin hexane) at −78° C. After stirring at −78° C. for 2 hours the reactionmixture was allowed to warm to −40° C. and a solution of 4.1 mL ofsulfuryl chloride in 75 mL of dry hexane was added within 15 minutes.Stirring was continued for 1 hour at −30° C. and subsequently foranother hour at 5° C. The mixture was poured on ice, extracted withdiethyl ether and the ether extract washed with cold water and brine,dried (sodium sulfate), and concentrated. The residue was purified bysilica chromatography (isohexane/ethyl acetate=4/1) yielding 0.62 g of4-(tetrahydropyran-4-yloxy)-benzenesulfonyl chloride as a colorless oil.

[0412] EI-MS: 276 (M⁺).

[0413] 58c.N-[1-(4-Amino-thieno[3,2-c]pyridin-2-ylmethyl)-2-(4-methyl-piperidin-1-yl)-2-oxo-ethyl]-4-(tetrahydropyran-4-yloxy)-benzenesulfonamide

[0414] 10 mL of a saturated solution of hydrogen chloride in diethylether were aided dropwise to a mixture of 210 mg of 30a in 15 mL of drydichloromethane and the reaction mixture was stirred for one hour atroom temperature. The solvent was removed in vacuo. 20 mL ofdichloromethane were added and subsequently removed in vacuo in order toget rid of traces of moisture. This procedure was repeated twice. Theresidue was dissolved in 15 mL of dichloromethane and 0.7 mL oftriethylamine and 207 mg of 4-(tetrahydropyran-4-yloxy)-benzenesulfonylchloride in 15 mL of dry dioxane were added dropwise. After stirring atroom temperature for 48 hours the solvents were evaporated in vacuo, theresidue redissolved in ethyl acetate, washed with water and brine, dried(sodium sulfate), and concentrated. Purification by silicachromatography (ethyl acetate/methanol=98/2, 96/4, 94/6, 92/8 v/v) gave100 mg of 58c.

[0415] M.p. 127-129° C., EI-MS: 558 (M⁺).

Example 59N-[1-(4-Amino-thieno[3,2-c]pyridin-2-ylmethyl)-2-(4-methyl-piperidin-1-yl)-2-oxo-ethyl]-4-(tetrahydropyran-4-yloxymethyl)-benzenesulfonamide(59c)

[0416] 59a. 4-(4-Bromo-benzyloxy)-tetrahydropyran

[0417] A mixture of 2.90 mL of 4-hydroxy-tetrahydropyran in 30 mL of dryN,N-dimethylformamide was added dropwise to a stirred suspension of 0.84g of sodium hydride (95%, dispersion in mineral oil) in 30 mL of dryN,N-dimethylformamide at 5° C. The resulting clear solution was allowedto come to room temperature and stirring was continued for 24 hours,followed by dropwise addition of 9.40 g of 4-bromo-benzylbromide in 50mL of dry N,N-dimethylformamide at 5° C. and stirring at roomtemperature for 24 hours. The mixture was poured into water andextracted with ethyl acetate. The combined extracts were washed withwater and brine, dried (sodium sulfate), and concentrated. The residuewas purified by silica chromatography (isohexane/ethyl acetate=9/1, 8/2,7/3, 6/4 v/v) yielding 5.40 g of 4-(4-bromo-benzyloxy)-tetrahydropyranas an oil. EI-MS: 270 (M⁺).

[0418] 59b. 4-(Tetrahydropyran-4-yloxymethyl)-benzenesulfonyl chloride

[0419] This compound was prepared from 0.68 g of4-(4-bromo-benzyloxy)-tetrahydropyran, 2 mL of n-butyllithium (1.6 N inhexane) and 0.81 mL of sulfuryl chloride using the procedure describedfor 58b. Yield: 0.70 g (oil), ¹H—NMR (D₆-DMSO): δ=1.41 (m, 2H), 1.86 (m,2H), 3.32 (m, 2H), 3.54 (m, 1H), 3.79 (m, 2H), 4.53 (s, 2H), 7.30 (d,2H), 7.61 (d, 2H).

[0420] 59c.N-[1-(4-Amino-thieno[3,2-c]pyridin-2-ylmethyl)-2-(4-methyl-piperidin-1-yl)-2-oxo-ethyl]-4-(tetrahydropyran-4-yloxymethyl)-benzenesulfonamide

[0421] This compound was prepared from 218 mg of4-(tetrahydropyran-4-yloxymethyl)-benzene-sulfonyl chloride and 210 mg(0.5 mmol) 30a using the procedure described for 58c. Yield: 100 mg,m.p. 124-128° C., (+)-FAB-MS: 573 (MH⁺).

Example 60N-[1-(4-Amino-thieno[3,2-c]pyridin-2-ylmethyl)-2-(4-methyl-piperidin-1-yl)-2-oxo-ethyl]-4-(2-methoxy-ethoxy)-benzenesulfonamide(60c)

[0422] 60a. (2-Methoxy-ethoxy)-benzene

[0423] A mixture of 4.70 g of phenol and 35 mL of dryN,N-dimethylformamide was added dropwise to a stirred suspension of 1.30g of sodium hydride (95%, dispersion in mineral oil) in 15 mL of dryN,N-dimethylformamide at 5° C. Stirring was continued for 2 hours at 5°C., followed by dropwise addition of 5.20 mL of 2-bromoethyl methylether (technical grade, 90%) at 5° C. The mixture was allowed to come toroom temperature and stirring was continued for 72 hours. The mixturewas poured into water and extracted with diethyl ether. The combinedextracts were washed with water and brine, dried (sodium sulfate) andconcentrated to give 5.90 g of (2-methoxy-ethoxy)-benzene as an oil.EI-MS: 159 (M⁺).

[0424] 60b. 4-(2-Methoxy-ethoxy)-benzenesulfonyl chloride

[0425] (2-Methoxy-ethoxy)-benzene (4.78 g) was dissolved in 50 mL ofchloroform and the solution was cooled to −10° C. Chlorosulfonic acid (4mL) was added dropwise while maintaining the temperature of the mixtureat −10° C. Stirring was continued for an additional hour at −10° C. Theprecipitated sulfonic acid was isolated by filtration and washed withcold chloroform and with cold isohexane (EI-MS: 232 (M⁺), m.p. 130-133°C.). 139 mg of the acid were dissolved in 5 mL of dichloromethane and125 mg of phosphorous pentachloride was added at 5° C. After stirringfor one hour at 5° C. complete reaction was indicated by TLC. Thevolatile components were removed in vacuo and the crude4-(2-methoxy-ethoxy)-benzenesulfonyl chloride was used in the next stepwithout further purification.

[0426] 60c.N-[1-(4-Amino-thieno[3,2-c]pyridin-2-ylmethyl)-2-(4-methyl-piperidin-1-yl-2-oxo-ethyl]-4-(2-methoxy-ethoxy)-benzenesulfonamide

[0427] This compound was prepared from 0.6 mmol4-(2-methoxy-ethoxy)-benzenesulfonyl chloride and 210 mg (0.5 mmol) of30a using the procedure described for 58c . Yield: 30 mg (whitecrystals), m.p. 210° C. (decomp.), EI-MS: 532 (M⁺).

Example 61N-[1-(4-Amino-thieno[3,2-c]pyridin-2-ylmethyl)-2-(4-methyl-piperidin-1-yl)-2-oxo-ethyl]-4-(2-methoxy-1-methoxymethyl-ethoxy)-benzenesulfonamide(61f)

[0428] 61a. 2-Methoxymethyl-oxirane

[0429] 8.1 mL of methanol and 1.3 g of tetrabutylammonium bromide weredissolved in 20 mL of 10 N sodium hydroxide, and 15.7 mL of2-chloromethyl-oxirane were added under vigorous stirring at such a ratethat the internal temperature did not exceed 30° C. Stirring wascontinued at room temperature for 24 hours, followed by extraction withdiethyl ether. The combined organic extracts were dried (sodium sulfate)and the solvents were removed. Distillation afforded 14.0 g of2-methoxymethyl-oxirane as a liquid. B.p. 20-25° C./12 Torr (reference:Davies, Alwyn G.; Hawari, Jalal A. -A.; Muggleton, Brenda; Tse,Man-Wing; J.Chem.Soc.Perkin Trans.2; 35, 1981; 1132-1137: b.p. 35° C.120 Torr).

[0430] 61b. 1,3-Dimethoxy-propan-2-ol

[0431] To a stirred solution of 13.2 g of 2-methoxymethyl-oxirane in 75mL of methanol was added 0.1 mL of boron trifluoride etherate whilestirring at 5° C. Stirring was continued for 24 hours at roomtemperature, followed by removal of the solvent. Distillation afforded7.0 g of 1,3-dimethoxy-propan-2-ol as a liquid. B.p. 75-80° C./12 Torr(reference: Gilchrist, Purves, J.Chem.Soc. 1925, 127, 2739, 2743: b.p.69.5-70.5/15 Torr); (+)-FAB-MS: 121 (MH⁺).

[0432] 61c. Methanesulfonic acid 2-methoxy-1-methoxymethyl-ethyl ester

[0433] 4.3 mL of methanesulfonyl chloride was added dropwise at 5° C. toa stirred solution of 6.0 g of 1,3-dimethoxy-propan-2-ol and 9.0 mL oftriethylamine in 60 mL of dichloromethane. Stirring was continued for 72hours at room temperature, followed by extraction with water. Thecombined organic extracts were dried (sodium sulfate) and concentratedand the residue purified by silica chromatography (isohexane/ethylacetate=9/1, 8/2, 7/3, 6/4 v/v) yielding 2.5 g of methanesulfonic acid2-methoxy-1-methoxymethyl-ethyl ester as an oil. EI-MS: 198 (M⁺).

[0434] 61d.(2-Methoxy-1-methoxymethyl-ethoxy)-benzene

[0435] A stirred mixture of 2.0 g of methanesulfonic acid2-methoxy-1-methoxymethyl-ethyl ester, 1.4 g of phenol and 2.8 g ofpowdered potassium carbonate in 50 mL of dry N,N-dimethylformamide washeated at 90° C. for 24 hours. After cooling, water was added at roomtemperature and the mixture extracted with diethyl ether. The combinedether extracts were washed with aqueous sodium hydroxide (2 N) andwater, dried (sodium sulfate) and concentrated. The crude product (1.0g) was used in the next step without further purification. EI-MS: 198(M⁺).

[0436] 61e. 4-(2-Methoxy-1-methoxymethyl-ethoxy)-benzenesulfonylchloride

[0437] (2-Methoxy-1-methoxymethyl-ethoxy)-benzene (1.0 g) was dissolvedin 10 mL of chloroform and the solution was cooled to −30° C.Chlorosulfonic acid (1 mL) was added dropwise while maintaining thetemperature of the mixture below −20° C. Stirring was continued insuccession for one hour at −20° C., for one hour at 5° C. and for 30minutes at room temperature. The mixture was poured on ice, the organiclayer washed with cold water, dried (sodium sulfate) and concentratedyielding 0.8 g of 4-(2-methoxy-1-methoxymethyl-ethoxy)-benzenesulfonylchloride as an oil. EI-MS: 294 (M⁺).

[0438] 61f.N-[1-(4-Amino-thieno[3,2-c]pyridin-2-ylmethyl)-2-(4-methyl-piperidin-1-yl)-2-oxo-ethyl]-4-(2-methoxy-1-methoxymethyl-ethoxy)-benzenesulfonamide

[0439] This compound was prepared from 0.75 mmol of4-(2-methoxy-1-methoxymethyl-ethoxy)-benzenesulfonyl chloride and 210 mg(0.5 mmol) of 30a using the procedure described for 58c.

[0440] Yield: 120 mg (oil), EI-MS: 576 (M⁺).

Example 62 Octane-1-sulfonic acid[1-(4-amino-thieno[3,2-c]pyridin-2-ylmethyl)-2-(4-methyl-piperidin-1-yl)-2-oxo-ethyl]-amide

[0441] This compound was prepared from 0.15 mL of octane sulfonylchloride and 210 mg of 30a using the procedure described for 58c. Yield:150 mg; m.p. 101-112° C., (+)-FAB-MS: 495 (M⁺).

Example 63 7-Methoxy-3,4-dihydro-1H-isoquinoline-2-sulfonic acid[1-(4-amino-thieno[3,2-c]pyridin-2-ylmethyl)-2-(4-methyl-piperidin-1-yl)-2-oxo-ethyl]-amide(63g)

[0442] 63a. Isoquinolin-7-ol

[0443] Aminoacetaldehyde dimethyl acetal (79 g) was added to a solutionof 106 g of 3-benzyloxy-benzaldehyde in 1100 mL of toluene The mixturewas refluxed for 6 hours using a Dean-Stark trap and subsequently cooledto 5° C. In a nitrogen atmosphere trifluoracetic acid anhydride (212 mL)and borontrifluoride etherate (185 mL) were added in succession at sucha rate, that the temperature of the mixture was kept below 10° C. Afterstirring for 5 days at room temperature the precipitated material wasseparated by filtration, washed with diethyl ether and dissolved in 750mL of water. The pH value was adjusted to 9 by adding concentratedaqueous ammonia. The precipitated product was separated by filtration,followed by washing with diethyl ether and drying in vacuo Yield: 53.2g; m.p. 210-218° C.; EI-MS: 145 (M⁺).

[0444] 63b. 1,2,3,4-Tetrahydro-isoquinolin-7-ol (hydroacetate)

[0445] 53.2 g of isoquinolin-7-ol were dissolved in 1000 mL of glacialacetic acid and hydrogenated for 48 hours at room temperature usingpre-hydrogenated platinum dioxide (3.5 g) as catalyst. Filtrationfollowed by concentration and addition of 50 mL of acetone gave a clearsolution. Addition of diethyl ether resulted in precipitation of pure1,2,3,4-tetrahydro-isoquinolin-7-ol-hydroacetate. Yield: 44.9 g;m.p.179-182° C.; EI-MS: 149 (M⁺).

[0446] 63c.7-Hydroxy-3,4-dihydro-1H-isoquinoline-2-carboxylic acidtert-butyl ester

[0447] A mixture of 17.1 g of di-tert-butyl-dicarbonate-in 170 mL ofdichloromethane was added dropwise to a stirred suspension of 16.4 g of1,2,3,4-tetrahydro-isoquinolin-7-ol-hydroacetate and 32.6 mL oftriethylamine in 164 mL of dichloromethane at 5° C. Stirring wascontinued at 5° C. for one hour, followed by evaporation of thevolatiles. The residue was redissolved in ethyl acetate and thissolution was washed in succession with aqueous acetic acid (1 N),saturated sodium hydrogen carbonate solution and brine and dried (sodiumsulfate). Removal of the solvent in vacuo gave pure7-hydroxy-3,4-dihydro-1H-isoquinoline-2-carboxylic acid tert-butyl esteras a solid. Yield: 17.5 g; m.p.140-142.5° C.; EI-MS: 249 (M⁺).

[0448] 63d. 7-Methoxy-3,4-dihydro-1H-isoquinoline-2-carboxylic acidtert-butyl ester

[0449] To a stirred mixture of 2.5 g of7-hydroxy-3,4-dihydro-1H-isoquinoline-2-carboxylic acid tert-butyl esterand 2.8 g of powdered potassium carbonate in 25 mL of dryN,N-dimethylformamide was added methyl iodide (0.94 mL) at 5° C.Stirring was continued for 24 hours at room temperature. Water was addedand the mixture was extracted with ethyl acetate. The combined organicextracts were washed with water and brine, dried (sodium sulfate) andthe solvent was removed in vacuo. The crude product (2.7 g; oil) wasused in the next step without further purification. EI-MS: 263 (M⁺).

[0450] 63e. 7-Methoxy-1,2,3,4-tetrahydro-isoquinoline

[0451] At 5° C., 50 mL of a saturated solution of hydrogen chloride indiethyl ether were added dropwise to a mixture of 2.3 g of7-methoxy-3,4-dihydro-1H-isoquinoline-2-carboxylic acid tert-butyl esterin 50 mL of diethyl ether and the reaction mixture stirred for 2 hoursat room temperature. The precipitated product (hydrochloride) wasseparated by filtration, washed with diethyl ether and dried in vacuo.Yield: 1.5 g (7.5 mmol); m.p. 215-218° C.; EI-MS: 163 (M⁺).

[0452] 63f. 7-Methoxy-3,4-dihydro-1H-isoquinoline-2-sulfonyl chloride

[0453] To a stirred, precooled solution (−40° C.) of 0.41 mL of sulfurylchloride in 10 mL of chloroform was added a mixture of 1.0 g of7-methoxy-1,2,3,4-tetrahydro-isoquinoline-hydrochloride and 2.1 mL oftriethylamine in 25 mL of chloroform at −40° C. Stirring was continuedat −30° C. for one hour and additionally at 5° C. for one hour. Themixture was poured on ice, the organic layer extracted with cold water,dried (sodium sulfate) and concentrated. The residue was redissolved indiethyl ether and chromatographed on silica gel (isohexane/ethylacetate=8/2 v/v) yielding 0.62 g of7-methoxy-3,4-dihydro-1H-isoquinoline-2-sulfonyl chloride as an oil.

[0454] EI-MS: 261 (M⁺).

[0455] 63g. 7-Methoxy-3,4-dihydro-1H-isoquinoline-2-sulfonic acid[1-(4-amino-thieno[3,2-c]pyridin-2-ylmethyl)-2-(4-methyl-piperidin-1-yl)-2-oxo-ethyl]-amide

[0456] This compound was prepared from 200 mg of7-methoxy-3,4-dihydro-1H-isoquinoline-2-sulfonyl chloride and 210 mg of30a using the procedure described for 58c. Yield: 110 mg; m.p. 175-176°C., (+)-FAB-MS: 544 (MH⁺).

Example 64 2,3-Dihydro-5H-benzo[f][1,4]oxazepine-4-sulfonic acid[1-(4-amino-thieno[3,2-c]pyridin-2-ylmethyl)-2-(4-methyl-piperidin-1-yl)-2-oxo-ethyl]-amide(64g)

[0457] 64a. 2-(Benzylimino-methyl)-phenol

[0458] A mixture of 5.0 mL of 2-hydroxy-benzaldehyde, 6.3 mL ofbenzylamine and 0.05 g of p-toluenesulphonic acid was refluxed intoluene for 2 hours using a Dean-Stark trap to remove liberated water.The volatiles were removed in vacuo yielding 10.1 g of crude2-(benzylimino-methyl)-phenol

[0459] 64b. 2-(Benzylamino-methyl)-phenol

[0460] Sodium borohydride (0.2 g) was added in small portions to 3 mL ofdry 2-propanol in a nitrogen atmosphere. To this mixture was addeddropwise a solution of 1.0 g of 2-(benzylimino-methyl)-phenol in 3 mL of2-propanol. After complete addition, stirring was continued for 1 h.Then water was added in order to decompose excess borohydride, and thevolatiles were removed in vacuo. The residue was redissolved in ethylacetate, washed with water, dried (sodium sulfate) and concentratedyielding 0.9 g of 2-(benzylamino-methyl)-phenol as an oil. EI-MS: 213(M⁺).

[0461] 64c. 4-Benzyl-4,5-dihydro-benzo[f][1,4]oxazepin-3-one

[0462] A mixture of 19.0 g of 2-(benzylamino-methyl)-phenol and 75 mL oftoluene was cooled to 0° C., and a solution of chloroacetyl chloride(7.8 mL) in 75 mL of toluene was slowly added while stirring. Stirringwas continued for 16 h. Dry N,N-dimethylformamide (60 mL) was added andthe solution was cooled to 0° C. In a nitrogen atmosphere 3:4 g ofsodium hydride (95%; dispersion in mineral oil) were added in smallportions and the mixture was allowed to stir at room temperature for 2h. Water was added and the aqueous layer was extracted with ethylacetate. The combined organic extracts were dried (sodium sulfate) andthe solvent was removed in vacuo yielding 12.4 g of4-benzyl-4,5-dihydro-benzo[f][1,4]oxazepin-3-one. M.p. 95-97° C.,

[0463] EI-MS: 253 (M⁺).

[0464] 64d. 4-Benzyl-2,3,4,5-tetrahydro-benzo[f][1,4]oxazepine

[0465] A solution of 2.5 g of4-benzyl-4,5-dihydro-benzo[f][1,4]oxazepin-3-one in 50 mL oftetrahydrofuran was added dropwise to a suspension of 1.14 g of lithiumaluminum hydride in 50 mL of tetrahydrofuran. The mixture was refluxedfor 5 hours and then allowed to cool down to room temperature. 2.2 mL ofsaturated aqueous sodium chloride solution was added in order todecompose excess lithium aluminum hydride. The precipitated solid wasremoved by filtration, and the filtrate was concentrated to give 2.2 gof pure 4-benzyl-2,3,4,5-tetrahydro-benzo[f][1,4]oxazepine as an oil.EI-MS: 239 (M⁺).

[0466] 64e. 2,3,4,5-Tetrahydro-benzo[f][1,4]oxazepine

[0467] 10.2 g of 4-benzyl-2,3,4,5-tetrahydro-benzo[f][1,4]oxazepine weredissolved in 100 mL of ethanol and hydrogenated for 5 hours at roomtemperature using 1.2 g of palladium (10% on carbon) as catalyst.Filtration followed by removal of the solvent in vacuo gave 6.1 g of2,3,4,5-tetrahydro-benzo[f][1,4]oxazepine as an oil. EI-MS: 149 (M⁺).

[0468] 64f. 2,3-Dihydro-5H-benzo[f][1,4]oxazepine-4-sulfonyl chloride

[0469] This compound was prepared from 1.5 g of2,3,4,5-tetrahydro-benzo[f][1,4]oxazepine, 0.81 mL of sulfuryl chlorideand 1.8 mL of ethyl diisopropyl amine using the procedure described for63f.Yield: 0.3 g (oil), EI-MS: 247 (M⁺).

[0470] 64g. 2,3-Dihydro-5H-benzo[f][1,4]oxazepine-4-sulfonic acid[1-(4-amino-thieno[3,2-c]pyridin-2-ylmethyl)-2-(4-methyl-piperidin-1-yl)-2-oxo-ethyl-amide

[0471] This compound was prepared from 200 mg of2,3-dihydro-5H-benzo[f][1,4]oxazepine-4-sulfonyl chloride and 210 mg of30a using the procedure described for 58c. Yield: 80 mg (oil),(+)-FAB-MS: 530 (MH⁺).

Example 65 7-Methoxy-2,3-dihydro-5H-benzo[f][1,4]oxazepine-4-sulfonicacid[1-(4-amino-thieno[3,2-c]pyridin-2-ylmethyl)-2-(4-methyl-piperidin-1-yl)-2-oxo-ethyl]-amide(65g)

[0472] 65a. 2-(Benzylimino-methyl)-4-methoxy-phenol

[0473] This compound was prepared from 2.7 g of2-hydroxy-5-methoxy-benzaldehyde, 2.3 mL of benzylamine and 0.03 g ofp-toluenesulphonic acid using the procedure described for 64a. Yield:4:3 g (oil).

[0474] 65b. 2-(Benzylamino-methyl)-4-methoxy-phenol

[0475] This compound was prepared from 1.0 g of2-(benzylimino-methyl)-4-methoxy-phenol and 0.18 g of sodium borohydrideusing the procedure described 64b. Yield: 1.0 g (oil), EI-MS: 243 (M⁺).

[0476] 65c. 4-Benzyl-7-methoxy-4,5-dihydro-benzo[f][1,4]oxazepin-3-one

[0477] A mixture of 2.43 g of 2-(benzylamino-methyl)-4-methoxy-phenoland 20 mL of toluene was cooled to 5° C. and a solution of chloroacetylchloride (0.88 mL) in 5 mL of toluene slowly added while stirring.Stirring was continued for 2 hours at room temperature. Drytetrahydrofuran (25 mL) was added and 3.4 g of sodium hydride (95%;dispersion in mineral oil) were added in small portions under a nitrogenatmosphere. The mixture was allowed to stir at room temperature for 24h. Water was added and the aqueous layer was extracted with ethylacetate. The combined organic extracts were dried (sodium sulfate) andthe solvent removed in vacuo yielding 2.0 g of4-benzyl-7-methoxy-4,5-dihydro-benzo[f][1,4]oxazepin-3-one as an oil.EI-MS: 283 (M⁺).

[0478] 65d. 4-Benzyl-7-methoxy-2,3,4,5-tetrahydro-benzo[f][1,4]oxazepine

[0479] This compound was prepared from 5.7 g of4-benzyl-7-methoxy-4,5-dihydro-benzo[f][1,4]-oxazepin-3-one and 2.3 g oflithium aluminum hydride using the procedure described for 64d.

[0480] Yield: 6.1 g (oil), EI-MS: 269 (M⁺).

[0481] 65e. 7-Methoxy-2,3,4,5-tetrahydro-benzo[f][1,4]oxazepine

[0482] This compound was prepared from 6.1 g of4-benzyl-7-methoxy-4,5-dihydro-benzo[f][1,4]-oxazepine and 1.0 g ofpalladium (10% on carbon) as catalyst using the procedure described for64e Yield: 2.7 g; m.p. 96-97° C., EI-MS: 179 (M⁺).

[0483] 65f. 7-Methoxy-2,3-dihydro-5H-benzo[f][1,4]oxazepine-4-sulfonylchloride

[0484] This compound was prepared from 1.4 g of7-methoxy-2,3,4,5-tetrahydro-benzo[f][1,4]-oxazepine, 0.91 mL ofsulfuryl chloride and 4.7 mL of triethylamine using the proceduredescribed for 63f. Yield: 0.23 g (oil), EI-MS: 277 (M⁺).

[0485] 65g. 7-Methoxy-2,3-dihydro-5H-benzo[f][1,4]oxazepine-4-sulfonicacid[1-(4-amino-thieno[3,2-c]pyridin-2-ylmethyl)-2-(4-methyl-piperidin-1-yl)-2-oxo-ethyl]-amide

[0486] This compound was prepared from 210 mg of7-methoxy-2,3-dihydro-5H-benzo[f][1,4]-oxazepine-4-sulfonyl chloride and210 mg of 30a using the procedure described for 58c. Yield: 70 mg (oil),(+)-FAB-MS: 560 (MH⁺).

Example 66(3S)-4-[[1-[(1-amino-6-isoquinolinyl)methyl]-2-oxo-2-(1-piperidinyl)ethyl]amino]-3-[[4-(3,5-dimethylisoxazolyl)sulfonyl]amino]-4-oxo-butanoicacid 1,1-dimethylethylester hydrochloride

[0487] The procedure described for 5c was used. Deprotection of 100 mgof 5a and coupling with 96 mg of(2S)-2-[[4-(3,5-dimethylisoxazolyl)sulfonyl]amino]butanedioic acid4-(1,1-dimethylethyl)ester (prepared from Asp(OtBu)-OH and3,5-dimethylisoxazole-4-sulfonyl chloride using the procedure describedfor 5b) yielded the title compound (66 mg) as a mixture of diastereomers(1:1). ¹H—NMR 400 MHz (CD₃OD) δ: 1.15-1.68 (6H, m), 1.40 (9H, s),2.20-2.62 (2H, m), 2.31 (3H, s), 2.33 and 2.37 (3H, 2× s), 2.96-3.56(6H, m), 4.10-4.14 (1H, m), 5.04-5.14 (1H, m), 7.16-7.23 (1H, m),7.53-7.78 (3H, m), 8.32-8.38 (1H, m).

Example 67(2S)-N¹-[1-[(1-amino-6-isoquinolinyl)methyl]-2-oxo-2-(1-piperidinyl)ethyl]-2-[[(4-methoxy-2,3,6-trimethylphenyl)sulfonyl]amino]-butanediamidehydrochloride

[0488] The procedure described for 5c was used. Deprotection of 100 mgof 5a and coupling with 105 mg of(2S)-4-amino-2-[[(4-methoxy-2,3,6-trimethylphenyl)sulfonyl]amino]-4-oxobutanoicacid (prepared from Asn-OH and 4-methoxy-2,3,6-trimethylsulfonylchloride(Mtr-chloride)using the procedure described for 5b) yielded the titlecompound (66 mg) as a mixture of diastereomers (1:1). ¹H—NMR 400 MHz(CD₃OD) δ: 1.20-1.68 (6H, m), 2.15 and 2.16 (3H, 2× s), 2.27-2.58 (2H,m), 2,57 (3H,s), 2.64 (3H, s), 2.93-3.57 (6H, m), 3.83 and 3.87 (3H, 2×s), 3.98-4.09 (1H, m), 5.07-5.19 (1H, m), 6.75 and 6.76 (1H, 2× s),7.22-7.27 (1H, m), 7.55-7.79 (3H, m), 8.33-8.38 (1H, m).

Example 68(3S)-4-[[1-[(1-amino-6-isoquinolinyl)methyl]-2-oxo-2-(4-morpholinyl)ethyl]amino]-3-[[(4-methoxy-2,3,6-trimethylphenyl)sulfonyl]amino]-4-oxo-butanoicacid 1,1-dimethylethylester hydrochloride

[0489] Protection of 0.23 g of amino acid 1i and subsequently couplingwith morpholine according to the procedure described for 5a yielded1,1-dimethylethyl[1-[(1-amino-6-isoquinolinyl)methyl]-2-oxo-2-(4-morpholinyl)ethylcarbamate.The procedure described for 5c was used for the deprotection of 90 mg of1,1-dimethylethyl[1-[(1-amino-6-isoquinolinyl)methyl]-2-oxo-2-(4-morpholinyl)ethylcarbamateand coupling with 96 mg of(2S)-2-[[(4-methoxy-2,3,6-trimethylphenyl)sulfonyl]amino]butanedioicacid 4-(1,1-dimethylethyl)ester (prepared from Asp(OtBu)-OH and(4-methoxy-2,3,6-trimethylphenyl)sulfonylchloride using the proceduredescribed for 5b) to yield the title compound (56 mg) as a mixture ofdiastereomers (1:1). ¹H—NMR 400 MHz (CD₃OD) δ: 1.29 and 1.33 (9H, 2× s),2.12 and 2.13 (3H, 2× s), 2.19-2.47 (2H, m), 2.52 and 2.53 (3H, 2× s),2.62 (3H, s), 2.95-3.65 (10H, m), 3.81 and 3.86 (3H, 2× s), 3.95-4.08(1H, m), 5.03-5.18 (1H, m), 6.73 and 6.75 (1H, 2× s), 7.18-7.23 (1H, m),7.53-7.78 (3H, m), 8.31-8.38 (1H, m).

Example 691-[3-(4-Aminothieno[3,2c]pyridin-2-yl)-2-[(1-oxo-2-propylpentyl)amino]-1-oxopropyl]-4-methylpiperidinehydrochloride

[0490] Compound 30a (53 mg) was dissolved in 1.5 mL of dichloromethaneand 1.5 mL of trifluoroacetic acid was added. After stirring at roomtemperature for 1.5 hours the reaction mixture was concentrated andcoevaporated with toluene. The residue was dissolved in 2 mL ofdichloromethane and 0.053 mL of triethylamine, cooled at 0° C. and 23 mgof 2-propylpentanoyl chloride was added. After stirring at roomtemperature for 1.5 h, dichloromethane and water were added. The organiclayer was separated, dried (magnesium sulfate) and concentrated.Purification by chromatography on silica gel (dichloromethane:methanol=95:5 (v/v)) yielded the free base. Treatment of this free basewith one equivalent hydrochloric acid and lyophilisation yieldedcompound 69 (22 mg). ¹H—NMR 400 MHz (CD₃OD) δ: 0.61-1.74 (22H, m), 2.182.27 (1H, m), 2.59-2.70 (1H, m), 2.99-3.10 (1H, m), 3.25-3.49 (2H, m),3.99-4.16 (1H, m), 4.41-4.51 (1H, m), 5.22-5.35 (1H, m), 7.37-7.42 (1H,m), 7.58-7.61 (2H, m).

Example 701-[3-4-Aminothieno[3,2c]pyridin-2-yl)-2-[[3-(4-chlorophenyl)-1-oxopropyl]amino]-1-oxopropyl]-4-methylpiperidinehydrochloride

[0491] Compound 30a (60 mg) was dissolved in 1.5 mL of dichloromethaneand 1.5 mL of trifluoroacetic acid was added. After stirring at roomtemperature for 1.5 h, the reaction mixture was concentrated andcoevaporated with toluene. The residue was dissolved in 2 mL ofdichloromethane and adjusted to pH 8 using N,N-diisopropylethylamine.This solution was added to an active ester solution that was prepared bydissolving 29 mg of 3-(4-chlorophenyl)propanoic acid, 21 mg ofhydroxybenztriazole and 33 mg of dicyclohexylcarbodiimide in 2.5 mL ofdichloromethane and stirring at room temperature for 15 min. After theaddition of the active ester solution the pH of the reaction mixture wasadjusted to 8 using N,N-diisopropylamine and stirred for 16 h at roomtemperature. The reaction mixture was filtered, water was added, and theorganic layer was separated. The aqueous layer was extracted withdichloromethane, the combined organic layers were dried (magnesiumsulfate) and concentrated. Purification by chromatography on silica gel(dichloromethane: methanol=9:1 (v/v)) yielded the free base. Treatmentof this free base with one equivalent hydrochloric acid andlyophilisation yielded the title compound (66 mg). ¹H—NMR 400 MHZ(CDCl₃) δ: 0.88 and 0.92 (3H, 2× d, J=6 Hz), 0.74-1.72 (5H, m),2.54-3.44 (8H, m), 3.81-3.92 (1H, m), 4.39-4.46 (1H, m), 5.17-5.23 (1H,m), 6.99-7.45 (7H, m), 7.82 (1H, br. s), 8.36 (2H, br.s).

Example 71 Solid-phase synthesis of compounds of formula (Ib) with n=0,X=S, R¹Y=R¹SO₂, R³=H (Table 71)

[0492] 71a.3-[4-aminothieno[3,2c]pyridin-2-yl]-2-[N-tert.butyloxycarbonyl]aminopropionic acid methyl ester

[0493] 9.3 g of compound 3h was coevaporated twice with dry methanol andsubsequently dissolved in 300 mL of methanol/triethylamine (9:1 v/v).7.5 g of di-tert.butyl carbonate was added and the reaction mixture wasstirred for 2 hours at room temperature. Three additional portions of1.5 g of di-tert.butyl carbonate were added over the next 5 hours. Thereaction mixture was evaporated and coevaporated twice with methanol.The residue was purified with silica gel chromatography with a gradientof methanol (0->8%) in dichloromethane containing 2 vol % oftriethylamine. This afforded 6.9 g of 71a. ¹H NMR 200 MHZ (CD₃OD) δ:1.41 (9H,s), 3.34 (2H,dd), 3.75 (3H,s), 4.46 (1H,dd), 7.07 (1H,d), 7.29(1H,s), 7.67 (1H,d).

[0494] 71b. 3-[4-(acetylamino)thieno[3,2c]pyridin-2-yl-2-[N-tertbutyloxycarbonyl]amino propionic acid methyl ester

[0495] 6.9 g of 71a was dissolved in 67 mL of pyridine and 1.8 mL ofacetic anhydride was added. After 2 hours at room temperature another1.8 mL of acetic anhydride was added and the solution was stirredovernight at room temperature. The pyridine was evaporated, and thecrude product was coevaporated three times with toluene. The residue waschromatographed on a silica gel column (ethyl acetate/methanol 98:2) togive 5.7 g of 71b. ¹H NMR 400 MHZ (CDCl₃) δ: 1.45 (9H,s), 2.32 (3H,s),3.45 (2H,dd), 3.80 (3H,s), 4.68 (1H,dd), 7.18 (1H,s), 7.52 (1H,d),8.11(1H, d).

[0496] 71c.3-[4-(acetylamino)thieno[3,2c]pyridin-2-yl]-2-[N-tert.butyloxycarbonyl]aminopropionic acid

[0497] 5.7 g of 71b was dissolved in 90 mL of dioxane/water (1:1 v/v)and 2N NaOH was added until pH 12. During the saponification was pHcontinuously adjusted to maintain pH 12. After 2 hours TLC(dichloromethane/methanol 9:1 v/v containing 0.5% triethylamine) showedcomplete conversion into the acid. The solution was neutralized withacetic acid and the mixture was concentrated to a small volume. Thesolution was acidified until pH 4 with acetic acid and extracted withethyl acetate. Between both layers a solid (sodium salt) appeared whichwas collected by filtration. The salt was added to adichloromethane/water mixture and acetic acid was added until pH 4.Again a solid (free acid) appeared between both layers. Filtration gave4.0 g of free acid 71c. ¹H NMR 200 MHZ (CD₃OD) δ: 1.40(9H,s), 2.28(3H,s), 3.43 (2H, dd), 4.44 (1H,dd), 7.34 (1H,s), 7.79 (1H,d),8.13(1H,d).

[0498] 71d. Derivatization of Kaiser oxime resin with acid 71c.

[0499] 2.0 g of 71c was coevaporated twice with dryN,N-dimethylformamide and subsequently dissolved in 50 mL ofdichloromethane/N,N-dimethylformamide (3:2 v/v). 0.88 g ofN-hydroxybenzotriazole was added and the resulting solution was added to3.5 g of Kaiser oxime resin (0.4 mmol/g). After the addition of 1.0 mLof diisopropylcarbodiimide, the suspension was shaken overnight at roomtemperature. The resin was filtered off and washed withdichloromethane/N,N-dimethylformamide (3:2 v/v) andN,N-dimethylformamide. Further washings were performed by alternateaddition of 2-propanol and dichloromethane (three times each). Unreactedoxime functions were capped by treatment of the resin with 35 mL aceticanhydride/N,N-diisopropylethylamine/N,N-dimethylformamide (3:1:12 v/v/v)for 30 minutes at room temperature. The resin was filtered off andwashed with N,N-dimethylformamide, 2-propanol and dichloromethane (threetimes each). The resin was dried in vacuo to give 4.2 g of 71d.

[0500] 71e.3-(4-aminothieno[3,2c]pyridin-2-yl)-2-[[(4-chlorophenyl)sulfonyl]amino]-N-[2-(3-fluorophenyl)ethyl)]-propanamide(compound of formula (Ib) with n=0, X=S, R¹Y=4-chlorophenylsulfonylR³=H, R⁷=3-fluorophenylethyl, R⁸=H.

[0501] 50 mg (20 μmol) of 71d was treated with 2 mL of 25 vol %trifluoroacetic acid in dichloromethane for 30 minutes at roomtemperature. The resin was filtered off and washed with dichloromethane2-propanol and dichloromethane. The resin was washed three times with 1mL dichloromethane/N,N-dimethylformamide (3:2 v/v) containing 80 μmolN,N-diisopropylethylamine and immediately reacted with 80 μmolp-chlorobenzenesulfonyl chloride in 1 mLdichloromethane/N,N-dimethylformamide (3:2 v/v) containing 80 μmolN,N-diisopropylethylamine. The suspension was shaken for 45 minutes atroom temperature. The resin was filtered off and washed withdichloromethane/N,N-dimethylformamide (3:2 v/v), followed by washingswith dichloromethane, 2-propanol and dichloromethane. The ninhydrin testrevealed complete conversion of the amine. The resin was suspended in 1mL of a 0.5 M solution of 3-fluorophenethylamine in distilledtetrahydrofuran and shaken for 16 hours at room temperature. The resinwas filtered off and washed with tetrahydrofuran and methanol. Thefiltrates were collected and concentrated to dryness. The residue wasdissolved in 1 mL ethylenediamine/ethanol (1:1 v/v) and shaken for 16hours at room temperature. The reaction mixture was evaporated todryness, dissolved in methanol/water (1:1 v/v) and applied to aDOWEX-NH₄ ⁺ column (4.5 mL) to remove excess of amine. The column waseluted with methanol/water (1:1 v/v). The TV positive fractions werepooled and evaporated to dryness yielding 9.5 mg of 71e.

Table 71 Solid-phase synthesis of compounds of formula (Ib) with n=0,X=S, R¹Y=R¹SO₂, R³=H

[0502] Using the procedure described for example 71e the sulfonylchlorides (R¹SO₂Cl) corresponding to R¹ in Table 71 were coupled toderivatized resin 71d. 50 mg portions of the resulting resins weretreated with amines of structure NHR⁷R⁸ as depicted in Table 71. Work-upof the samples was performed as described for 71e.

[0503] All compounds were characterized by reversed phase liquidchromatography on a Supelcosil LC-18-DB column using followingconditions: Flow: 1.0 ml/min; Buffers A: water, B: acetonitrile/water(9:1 v/v), C: 0.5M phosphate buffer pH=2.1; Gradient 1:0→30 min 65%A-15% B-20% C→25% A-55% B-20% C. UV-detection at 210 nm. Retention timesare given in minutes in Table 71. TABLE 71 RP-HPLC retention times forcompounds of example 71

R¹SO₂ R⁷NR⁸

30.6 23.0 26.7 23.1 28.9

29.9 23.9 26.6 22.4 28.3

30.4 24.3 27.1 24.2 29.0

29.2 23.9 26.2 22.8 28.0

32.2 28.9 24.4 30.5

26.6 19.1 22.8 18.0 24.7

13.6  6.3  8.5  3.5 11.8

33.8 28.1 30.7 27.3 32.1

Example 72 Solid-phase synthesis of compounds of formula (Ib) with n=0,X=S, R¹Y=R¹C(O), R³=H (Table 72)

[0504] 72a3-(4-aminothieno[3,2c]pyridin-2-yl)-N-[methyl]-N-[phenylmethyl)]-2-[[(2-pyridinyl)-carbonyl]amino]-propanamide(compound of formula (Ib) with n=0, X=S, R¹Y=2-pyridine-carboxyl, R³=H,R⁷=benzyl, R⁸=methyl)

[0505] 50 mg (20 μmol) of 71d was treated with 2 mL of 25 vol %trifluoroacetic acid in dichloromethane for 30 minutes at roomtemperature. The resin was filtered off and washed with dichloromethane,2-propanol and dichloromethane. The resin was washed three times with 1mL dichloromethane/N,N-dimethylformamide (3.2 v/v) containing 80 μmolN,N-diisopropylethylamine and immediately reacted with 80 μmol2-pyridinecarboxylic acid in 1 ml dichloromethane/N,N-dimethylformamide(3:2 v/v) containing 80 μmol of N,N-diisopropylethylamine and 80 μmol of(O-(benzotriazol-1-yl))-1,1,3,3-tetramethyl uronium tetrafluoroborate.The suspension was shaken for 45 minutes at room temperature. The resinwas filtered off and washed with dichloromethane/N,N-dimethylformamide(3:2 v/v), followed by washings with dichloromethane, 2-propanol anddichloromethane. The ninhydrin test revealed complete conversion of theamine.

[0506] The resin was suspended in 1mL of a 0.5 M solution ofmethylbenzylamine in distilled tetrahydrofliran and shaken for 16 hoursat room temperature. The resin was filtered off and washed withtetrahydrofuran and methanol. The filtrates were collected andconcentrated to dryness. Further processing of the sample was performedas described for 71e, yielding 8.0 mg of compound 72a.

Table 72 Solid-phase synthesis of compounds of formula (Ib) with n=0,X=S, R¹Y=R¹C(O), R³=H

[0507] Using the procedure described for example 72a the carboxylicacids (R¹C(O)OH) corresponding to R¹ in Table 72 were coupled toderivatized resin 71d. 50 mg portions of the resulting resins weretreated with amines of structure NHR⁷R⁸ as depicted in Table 72. Work-upof the samples was performed as described for 71e.

[0508] All compounds were characterized by reversed phase liquidchromatograpy using the conditions described for compounds 71f.Retention times are given in minutes in Table 72. Standard gradient 1was run, times marked with an asterisk were determined after applyingthe following gradient: 0→30 min 75% A-5% B-20% C→35% A-45% B-20% C.TABLE 72 RP-HPLC retention times for example 72

R¹C(O)              R⁷NR⁸

21.0 31.9 23.4 28.3/29.0 24.6 16.6

22.4 32.0 23.0 28.6/29.5 24.7 17.1

22.9 31.8 24.2 28.9/29.7 18.3

22.3 30.8 23.4 28.5/29.6 18.9

24.3 30.2/31.2 25.9 19.3

17.3 28.3 18.7 24.1/25.3 20.7 12.4

14.3* 22.7* 15.9* 20.5/21.3* 18.4* 10.7*

27.9 36.9 28.2 33.6/34.6 30.3 24.1

Example 73N-(carboxymethyl)-D-cyclohexylalanyl-N-[(4-aminothieno[3,2c]pyridin-2-yl)methyl]-(N-cyclopentyl-glycin)amide(N—(HOOCCH₂)-D-Cha-N-cyclopentyl-Gly-Atp) (73j)

[0509] 73a. N-[2-(azidomethyl)thieno[3,2c]pyridin-4-yl] benzamide

[0510] The experimental procedure described in J. Org. Chem. 58, 5886(1993) was slightly modified. To 1.0 g ofN-[2-(hydroxymethyl)thieno[3,2c]pyridin-4-yl]benzamide (3e) was addedtoluene and evaporated under reduced pressure to remove traces ofmoisture. To the residue was added 5 mL of toluene and 5 mL of dioxane,the solution was cooled at 0° C. and 1.14 mL of diphenylphosphoryl azide(DDPA) and 0.79 mL 1,8-diazabicyclo(5.4.0)undec-7ene (DBU) were addedAfter stirring this suspension for 68 hours at room temperature thereaction was not completed and an additional 0.38 mL of DPPA and 0.27 mLof DBU were added. After 24 hours the reaction mixture was poured intowater (pH 7) and extracted with ethyl acetate. The organic layer wasdried (magnesium sulfate) and concentrated. Purification on silica gel(toluene/ethyl acetate: 3/1 (v/v)) gave 0.98 g azide. ¹H—NMR 200 MHz(CDCl₃) δ: 4.59 (2H, s), 7.38-7.59 (5H, m), 8.01-8.09 (3H, m).

[0511] 73b N-[2-(aminomethyl)thieno[3,2c]pyridin-4-yl]benzamide

[0512] Through a solution of 1.07 g ofN-[2-(azidomethyl)thieno[3,2c]pyridin-4-yl]benzamide in 50 mL of ethanolcontaining 0.6 g of 5% palladium/calcium carbonate was bubbled hydrogenfor 40 hours. Filtration and column chromatography on silica gel(dichloromethane/methanol=9/1 (v/v)) yielded 0.79 g of the titlecompound. ¹H—NMR 200 z (CDCl₃) δ: 4.18 (2H, s), 7.35-7.63 (5H, m),7.99-8.12 (3H, m).

[0513] 73c. 4-amino-2-(aminomethyl)thieno[3,2c]pyridine hydrochloride

[0514] A solution of 0.33 g ofN-[2-(aminomethyl)thieno[3,2c]pyridin-4-yl]benzamide in 40 mL of 4Nhydrochloric acid and 20 mL of acetic acid was refluxed for 16 hours.The reaction mixture was washed with diethylether to remove benzoic acidand the aqueous solution was concentrated under reduced pressure to give0.256 g of 4-amino-2-(aminomethyl) thieno[3,2c]pyridine hydrochloride.¹H—NMR 200 MHz (D₂O) δ: 4.57 (2H, s), 7.43 (1H, d, J=7 Hz), 7.63 (1H, d,J=7 Hz), 7.83 (1H, s).

[0515] 73d. N-Cyclopentyl-Gly-OMe

[0516] Cyclopentanone (15.6 g) was added to a solution of H-Gly-OMe HCl(23.2 g) in 200 mL of methanol. The mixture was stirred for 15 minutesand sodium cyanoborohydride (7 g) was added. The pH was adjusted to 6.The reaction mixture was stirred for 16 hours at room temperature. Tocomplete the reaction cyclopentanone (1 g) was added and stirring wascontinued.The reaction was monitored on TLC. When all the startingmaterial had disappeared, the mixture was acidified to pH 2 and wasstirred for 30 minutes. The solvent was removed and the residue wasdiluted with water. The solution was washed with ether, the pH adjustedto 12 with 6N sodium hydroxide and extracted with dichloromethane. Thecombined organic layers were washed with a saturated sodium chloridesolution, dried on sodium sulfate and evaporated in vacuo to yield 16 gof an oil.

[0517] Rf=0.46 in ethyl acetate/pyridine/acetic acid/water 63/20/6/11(v/v/v/v) on silica.

[0518] 73e. N-(t-butyloxycarbonylmethyl)-D-Cha-OMe

[0519] t-Butyl bromo acetate (17 g) was added to a stirred solutionH-D-Cha-OMe HCl (26 g) in 300 mL of acetonitrile. The pH of the mixturewas adjusted to 8.5 with N,N-diisopropylethylamine. The mixture wasstirred for 16 hours at room temperature and evaporated in vacuo. Theresidue was dissolved in dichloromethane and the solution was washedwith water, dried on sodium sulfate and evaporated in vacuo.Chromatography over silica gel in hexane/ethyl acetate 9/1 (v/v) gave 20g of N-(t-butyloxycarbonylmethyl)-D-Cha-OMe.

[0520] Rf=0.46 in ethyl acetate/pyridine/acetic acid/water 63/20/6/11(v/v/v/v) on silica.

[0521] 73f. N-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-OMe

[0522] The pH of a solution of N-(t-butyloxycarbonylmethyl)-D-Cha-OMe(20 g) and di-t-butyl dicarbonate (17 g) was adjusted to 8.5 withN,N-diisopropylethylamine. The mixture was stirred for 16 hours at roomtemperature. The solvent was removed in vacuo. Dichloromethane and waterwere added to the residue. The organic layer was separated, washed withcold 1N hydrochloric acid, water, 5% sodium hydrogen carbonate andwater. The organic layer was dried on sodium sulfate and the filtratewas evaporated to an amorphous solid ofN-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-OMe with a yield of 28 g.

[0523] Rf=0.60 in ethyl acetate/pyridine/acetic acid/water 252/20/6/11(v/v/v/v) on silica.

[0524] 73d. N-Cyclopentyl-Gly-OMe

[0525] Cyclopentanone (15.6 g) was added to a solution of H-Gly-OMe HCl(23.2 g) in 200 mL of methanol. The mixture was stirred for 15 minutesand sodium cyanoborohydride (7 g) was added. The pH was adjusted to 6.The reaction mixture was stirred for 16 hours at room temperature. Tocomplete the reaction cyclopentanone (1 g) was added and stirring wascontinued.The reaction was monitored on TLC. When all the startingmaterial had disappeared, the mixture was acidified to pH 2 and wasstirred for 30 minutes. The solvent was removed and the residue wasdiluted with water. The solution was washed with ether, the pH adjustedto 12 with 6N sodium hydroxide and extracted with dichloromethane. Thecombined organic layers were washed with a saturated sodium chloridesolution, dried on sodium sulfate and evaporated in vacuo to yield 16 gof an oil.

[0526] Rf=0.46 in ethyl acetate/pyridine/acetic acid/water 63/20/6/11(v/v/v/v) on silica.

[0527] 73e. N-(t-butyloxycarbonylmethyl)-D-Cha-OMe

[0528] t-Butyl bromo acetate (17 g) was added to a stirred solutionH-D-Cha-OMe HCl (26 g) in 300 mL of acetonitrile. The pH of the mixturewas adjusted to 8.5 with N,N-diisopropylethylamine. The mixture wasstirred for 16 hours at room temperature and evaporated in vacuo. Theresidue was dissolved in dichloromethane and the solution was washedwith water, dried on sodium sulfate and evaporated in vacuo.Chromatography over silica gel in hexane/ethyl acetate 9/1 (v/v) gave 20g of N-(t-butyloxycarbonylmethyl)-D-Cha-OMe.

[0529] Rf=0.46 in ethyl acetate/pyridine/acetic acid/water 63/20/6/11(v/v/v/v) on silica.

[0530] 73f. N-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-OMe

[0531] The pH of a solution of N-(t-butyloxycarbonylmethyl)-D-Cha-OMe(20 g) and di-t-butyl dicarbonate (17 g) was adjusted to 8.5 withN,N-diisopropylethylamine. The mixture was stirred for 16 hours at roomtemperature The solvent was removed in vacuo. Dichloromethane and waterwere added to the residue. The organic layer was separated, washed withcold 1N hydrochloric acid, water, 5% sodium hydrogen carbonate andwater. The organic layer was dried on sodium sulfate and the filtratewas evaporated to an amorphous solid ofN-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-OMe with a yield of 28 g.

[0532] Rf=0.60 in ethyl acetate/pyridine/acetic acid/water 252/20/6/11(v/v/v/v) on silica.

[0533] 73g. N-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-OH

[0534] A solution of N-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-OMe (28 g)in 420 mL of dioxane:water 9/1 (v/v) was treated with sufficient 1Nsodium hydroxide to keep the pH at 13 for 90 minutes at roomtemperature. After acidification, the mixture was poured into water andwas extracted with dichloromethane. The organic layer was washed withwater and was dried on sodium sulfate. The filtrate was evaporated andyielded 24 g of N-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-OH.

[0535] Rf=0.23 in dichloromethane/methanol 9/1 (v/v) on silica

[0536] 73h.N-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-N-cyclopentyl-Gly-OMe

[0537] N-cyclopentyl-Gly-OMe (10.2 g) and2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate(TBTU, 21.2 g) were added to a solution ofN-Boc-N-(t-butyloxy-carbonylmethyl)-D-Cha-OH (24 g) in 300 mL ofN,N-dimethyl formamide.

[0538] The pH of the mixture was adjusted to 8.5. The mixture wasstirred overnight at room temperature and was concentrated byevaporation. Water and ethyl acetate were added to the residue. Theorganic layer was separated and washed with 1N hydrochloric acid, water,5% sodium hydrogen carbonate and water and dried over sodium sulfate.The filtrate was evaporated and the residue was chromatographed onsilica gel in hexane/ethyl acetate 8/2 (v/v) as eluent. The fractionscontainingN-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-N-cyclopentyl-Gly-OMe werepooled and evaporated. Yield. 17 g.

[0539] Rf=0.57 in hexane/ethyl acetate 713 (v/v) on silica.

[0540] 73i.N-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-N-cyclopentyl-Gly-OH

[0541] N-Boc-N-(t-butyloxycarbonylmethyl)-Cha-N-cyclopentyl-Gly-OMe (17g) was saponified in a mixture of dioxane/water 1/1 (v/v, 150 mL) anddiluted sodium hydroxide and yielded 15 g of an amorphous solid.Chromatography over silica gel with dichloromethane/methanol 95/5 (v/v)as eluent gave 13 g ofN-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-N-cyclopentyl-Gly-OH.

[0542] Rf=0.30 in dichloromethane/methanol 9/1 (v/v) on silica.

[0543] 73j. N—(HOOCCH2)-D-Cha-N-cyclopentyl-Gly-Atp

[0544] To a solution of 91 mg of 4-amino-2-(aminomethyl)thieno[3,2c]pyridine . hydrochloride and 182 mg ofN-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-N-cyclopentyl-Gly-OH in 2.5 mLof N,N-dimethylformamide was added 125 mg2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate(TBTU) and the pH was adjusted to 8 using N,N-diisopropylethylamine. ThepH was maintained at 8 during the reaction. After 16 hours the reactionmixture was concentrated under reduced pressure. Dichloromethane wasadded, washed with an aqueous 5% sodium hydrogencarbonate solution,dried (magnesium sulfate) and concentrated. Purification on silica gel(dichloromethane/methanol: 95/5 (v/v)) gave 164 mg ofN-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-N-cyclopentyl-Gly-Atp as anoil. 142 mg of this oil was dissolved in 2.5 mL of dichloromethane and2.5 mL of trifluoroacetic acid and stirred at room temperature. After 16hours the reaction mixture was concentrated, dissolved in 0.2 Mhydrochloric acid and washed with ether. The aqueous solution wasconcentrated under reduced pressure. HPLC purification on a preparativeDelta-pack C-18 column using a gradient elution system of 20% A/70%B/10% C to 20% A/20% B/60% C (A=0.5M sodium dihydrogenphosphate+phosphoric acid (pH=2.1); B=water; C=cyanomethane/water 3/2(v/v) over 45 min, at a flow rate of 80 mL/min. Yield: 100 mg. ¹H—NMR400 MHz (D₂O) δ: 0.43-1.84 (21H, m), 3.40-3.67 (2H, m), 3.84-4.13 (3H,m), 4.46-4.7 (3H, m), 7.21-7.26 (1H, m), 7.39-7.48 (2H, m).

Example 74N-(carboxymethyl)-D-cyclohexylalanyl-[N-(1-amino-6-isoquinolinyl)methyl]-L-azetidin-2-carboxamidetrifluoroacetate (N—(HOOCCH₂)-D-Cha-L-Azt-6-Aig TFA)(74f)

[0545] 74a. N-[6-(azidomethyl)-1-isoquinolinyl]-benzamide

[0546] To 6.3 g of N-[6-(hydroxymethyl)-1-isoquinolinyl] benzamide (1g)in 40 mL of toluene and 40 mL of dioxane at 0° C. 7.8 mL ofdiphenylphosphoryl azide (DDPA) and 5.4 mL of1,8-diazabicyclo(5.4.0)undec-7ene (DBU) were added. After 24 hours thereaction mixture was poured into water (pH 7) and extracted with ethylacetate. The organic layer was dried (sodium sulfate) and concentrated.Trituration with diethyl ether gave 6.4 g of the title compound. Mp.125-129° C. MS (m/e)=303.

[0547]74b. N-6-(aminomethyl)-1-isoquinolinyl]benzamide

[0548] A solution of 6.4 g ofN-[6-(azidomethyl)-1-isoquinolinyl]benzamide in 300 mL of ethanol and 60mL of N,N-dimethylformamide containing 3.8 g of 5% palladium/calciumcarbonate was hydrogenated for 20 hours. Filtration and concentration i.vac. yielded the title compound as an oil. MS (m/e)=277.

[0549] 74c. 1-amino-6-(aminomethyl)isoquinoline hydrochloride

[0550] A solution of 5.8 g ofN-[6-(aminomethyl)-1-isoquinolinyl]benzamide in 300 mL of 4Nhydrochloric acid and 150 mL of acetic acid was refluxed for 6 hours.The reaction mixture was washed with diethylether to remove benzoic acidand the aqueous solution was concentrated under reduced pressure to give3.0 g of the title compound. MS (m/e)=173.

[0551] 74d.(N-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-(L)-azetidine-2-carboxylicacid

[0552] 0.71 mL (5.43 mmol) Isobutyl chloroformate was added dropwise toa cooled (−15-−20° C.) solution of 1.90 g (4.94 mmol)N-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-OH and 0.6 mL (5.43 mmol)N-methyl morpholine in 15 mL dichloromethane. After 45 min, 0.50 g (4.94mmol) (L)-azetidine-2-carboxylic acid (L-H-Azt-OH) was added. After 1 h,the organic phase was extracted with water, the organic layer was dried(sodium sulfate), filtered, and the solvent was removed i.vac. to yield2.3 g (quant.) of the title compound as a nearly colorless oil. FAB-MS:m/e=469 (M+H⁺).

[0553] 74e. N-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-L-Azt-6-Aiq

[0554] 0.45 g (1.28 mmol)Chloro-N,N,N′,N′-bis(pentamethylene)-formamidinium hexafluorophosphatewas added to a cooled (ice bath) solution of 0.60 g (1.28 mmol)N-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-L-Azt-OH (74d) and 0.23 g (1.28mmol) 1-amino-6-(aminomethyl)isoquinoline hydrochloride (74c) in 5 mLdichloromethane, followed by the addtion of 0.68 mL (3.83 mmol)N,N-diisopropylethylamine. After 2 h at room temperature, the solutionwas extracted with water, the organic layer was dried (sodium sulfate),filtered, and the solvent was removed i. vac. The residue was filteredover silica gel using ethyl acetate: methanol (15:1) as an eluent. Thesolvent was removed i vac. to yieldN-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-L-Azt-6-Aiq as an amorphoussolid. FAB-MS m/e 623.

[0555] 74f. N—(HOOCCH₂)-D-Cha-L-Azt-6-Aiq TFA 3.0 mL Trifluoroaceticacid was added to a solution of 200 mg (0.32 mmol)N-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-L-Azt-6-Aiq in 3 mLdichloromethane. After 3 h, the solvent was removed i. vac., the residuewas dissolved in methanol, filtered, and the solvent was removed i. vac.to yield 90 mg of an amorphous residue. The residue was purified bycolumn chromatography (reversed phase RP18 select B;methanol:water=55:45) to yield the title compound as an amorphous solid.FAB-MS: m/e=467.

Example 75N-(carboxymethyl)-D-cyclohexyalanyl-N-[(4-amino-thieno[3,2c]pyridin-2-yl)-methyl]-L-azetidin-2-carboxamidetrifluoroacetate (N—(HOOCCH₂)-D-Cha-L-Azt-Atp TFA)(75b)

[0556] 75a. N-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-L-Azt-Atp

[0557] 0.45 g (1.28 mmol)Chloro-N,N,N′,N′-bis(pentamethylene)-formamidinium hexafluorophosphatewas added to a cooled (ice bath) solution of 0.60 g (1.28 mmol)N-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-L-Azt-OH (74d) and 0.28 g (1.28mmol) 4-amino-2-(aminomethyl)thieno[3,2c]pyridine hydrochloride (73c) in5 mL dichloromethane, followed by the addition of 0.68 mL (3.83 mmol)N,N-diisopropylethylamine. After 15 min at 0° C. and 2 h at roomtemperature, the solution was filtered, the filtrate was extracted withwater, the organic layer was dried (sodium sulfate), filtered, and thesolvent was removed i. vac The residue was filtered over silica gelusing ethyl acetate:methanol (15:1) as an eluent. The solvent wasremoved in vacuo to yieldN-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-L-Azt-Atp as are amorphoussolid. FAB-MS m/e=629.

[0558] 75b. N—(HOOCCH₂)-D-Cha-L-Azt-Atp TFA)

[0559] 2.0 mL Trifluoroacetic acid was added to a solution of 85 mg(0.13 mmol) N-Boc-N-(butyloxycarbonylmethyl)-D-Cha-L-Azt-Atp in 2 mLdichloromethane. After 3 h, the solvent was removed in vacuo, theresidue was dissolved in methanol, filtered, and the solvent was removedin vacuo to yield 100 mg of an amorphous residue. The residue waspurified by column chromatography (reversed phase RP18 select B;methanol:water=60:40). The solvent was removed in vacuo, the residue wasdissolved in methanol, toluene was added, the solvent was removed invacuo, the residue was triturated with methanol, filtered, and thesolvent was removed in vacuo to yield the title compound as an amorphoussolid. FAB-MS: m/e=473.

Example 76N-Methyl-D-phenylalanyl-N-[(1-amino-6-isoquinolinyl)methyl]-L-prolinamidetrifluoroacetate (N-Me-D-Phe-Pro-6-Aiq TFA) (76b)

[0560] 76a.N-Boc-N-Me-D-Phe-Pro-6-Aiq

[0561] In the manner described in example 74eN-Boc-N-methyl-D-phenylalanyl-N-[(1-amino-6-isoquinolinyl)-methyl]L-prolineamide (N-Boc-N-Me-D-Phe-Pro-6-Aiq) was obtained by the reactionof N-Boc-N-methyl-D-phenylalanyl-L-proline (N-Boc-N-Me-D-Phe-Pro-OH)with 1-amino-6-(aminomethyl)isoquinoline hydrochloride (74c). Amorphoussolid FAB-MS m/e=531.

[0562] 76b.N-Me-D-Phe-Pro-6-Aiq TFA

[0563] The Boc-group of N-Boc-N-Me-D-Phe-Pro-6-Aiqwas removed asdescribed in example 74f, and the title compound was obtained as asolid, which decomposed on melting at 72° C. FAB-MS: m/e=431.

Example 77N-(carboxymethyl)-D-phenylalanyl-N-[(1-amino-6-isoquinolinyl)methyl]L-prolinamidetrifluoroacetate (N—(HOOCCH₂)D-Phe-Pro-6-Aiq TFA) (77f)

[0564] 77a. N-t-butyloxycarbonylmethyl)-D-Phe-OMe

[0565] 15 mL Hünig-base was added to a suspension of 100 gD-phenylalanine-methylester (D-H-Phe-OMe) and 5 mL t-butyl bromoacetatein 100 mL dichoromethane (pH=8). The suspension slowly converted into asolution, and after 20 hours at room temperature, it was extracted withwater, the organic layer was separated, dried (sodium sulfate),filtered, and concentrated i. vac. The residue was filtered (silica gel,elution with heptane/ethyl acetate=2:1). Concentration in vacuo yielded6.4 g of the tilte compound as an oil. MS (m/e)=293.

[0566] 77b. N-Boc-N-(t-butyloxycarbonylmethyl)-D-Phe-OMe

[0567] 25 mL Hünig-base was added to a solution of 16.7 gN-(t-butyloxycarbonylmethyl)-D-Phe-OMe and 14.8 g di-t-butyl dicarbonatein 150 mL N, N-dimethylformamide (pH=8). After 60 h at room temperature,the N, N-dimethylformamide was removed, the residue dissolved in ethylacetate, extracted with water, dried (sodium sulfate), filtered, andconcentrated in vacuo to yield the title compound as a yellow oil. MS(m/e)=393.

[0568] 77c. N-Boc-N-(t-butyloxycarbonylmethyl)-D-Phe-OH

[0569] 22.4 g N-Boc-N-(t-butyloxycarbonylmethyl)-D-Phe-OMe and 23 gsodium hydroxide were dissolved in 180 mL dioxane and 35 mL water. After26 hours at room temperature, the solvent was removed in vacuo, theresidue was dissolved in diethyl ether and extracted with water, theaqueous layer was acidified with conc hydrochloric acid and extractedwith ether. The organic layer was dried (sodium sulfate), filtered, andconcentrated in vacuo to yield 18.4 g of the title compound as a yellowoil. MS (m/e)=379.

[0570] 77d. N-Boc-N-(t-butyloxycarbonylmethyl)-D-Phe-Pro-OH

[0571] was prepared analogously to 74d usingN-Boc-N-(t-butyloxycarbonylmethyl)-D-Phe-OH and proline and isolated in79% yield as a yellow oil. MS (m/z) 476.

[0572] 77e. N-Boc-N-(t-butyloxycarbonylmethyl)-D-Phe-Pro-6-Aiq

[0573] In the manner described in example 74e,N-Boc-N-(t-butyloxycarbonylmethyl)-D-Phe-Pro-6-Aiq was obtained by thereaction of N-Boc-N-(t-butyloxycarbonylmethyl)-D-Phe-Pro-OH with1-amino-6-(aminomethyl)isoquinoline hydrochloride (74c). Amorphoussolid. FAB-MS: m/e=631.

[0574] 77f. N—(HOOCCH₂)-D-Phe-Pro-6-Aiq TFA

[0575] The Boc-group and t-butyl ester ofN-Boc-N-(t-butyloxycarbonylmethyl)-D-Phe-Pro-6-Aiq were removed asdescribed in example 74f, and the title compound was obtained as anamorphous solid. FAB-MS: m/e=475.

Example 78N-(carboxymethyl)-D-cyclohexylalanyl-N-[1-amino-6-isoquinolinyl)methyl]-(N-cyclopentyl-glycin)-amidetrifluoroacetate (N—(HOOCCH₂)-D-Cha-N-cyclopentyl-Gly-6-Aiq TFA) (78b)

[0576] 78a.N-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-N-cyclopentyl-Gly-6-Aiq

[0577] In the manner described in example 74e,N-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-N-cyclopentyl-Gly-6-Aiq wasobtained by the reaction ofN-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-N-cyclopentyl-Gly-OH (73i) with1-amino-6-(aminomethyl)isoquinoline hydrochloride (74c). Amorphoussolid. FAB-MS: m/e=665.

[0578] 78b. N—(HOOCCH₂)-D-Cha-N-cyclopentyl-Gly-6-Aiq TFA

[0579] The Boc-group and the t-butyl group were removed as described inexample 74f; and the title compound was obtained as an amorphous solid.FAB-MS: m/e=509.

Example 79N-(carboxymethyl)-D-cyclohexylalanyl-N-[(1-amino-6-isoquinolinyl)methyl]-(N-cyclohexyl-glycin)-amidetrifluoroacetate (N—(HOOCCH₂)-D-Cha-N-cyclohexyl-Gly-6-Aiq TFA)(79e)

[0580] 79a. N-Cyclohexyl-Gly-OMe

[0581] was prepared from cyclohexanone (36.9 g) and H-Gly-OMe HCl (46.4g) in 400 mL of methanol analogously to 73d to give 23.4 g of an oil. MS(m/e)=171 [M+].

[0582] 79b.N-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-N-cyclohexyl-Gly-OMe

[0583] was prepared from N-cyclohexyl-Gly-OMe andN-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-OH analogously to 73h.

[0584] 79c. N-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-N-cyclohexyl-Gly-OH

[0585] was prepared fromN-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-(N-cyclohexyl)-Gly-OMeanalogously to 73I.

[0586] 79d.N-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-N-cyclohexyl-Gly-6-Aiq

[0587] In the manner described in example 74e,N-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-N-cyclohexyl-Gly-6-Aiq wasobtained by the reaction ofN-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-N-cyclohexyl-Gly-OH with1-amino-6-(aminomethyl)isoquinoline hydrochloride (74c). Amorphoussolid. FAB-MS: m/e=679.

[0588] 79e. N—(HOOCCH₂)-D-Cha-(N-cyclohexyl)Gly-6-Aiq TFA

[0589] The Boc-group and the t-butyl group were removed as described inexample 74f, and the title compound was obtained as an amorphous solid.Mp. 99° C. (dec.); FAB-MS: m/e=523.

Example 80N-(carboxymethyl)-D-cyclohexylalanyl)-N-[(4-aminothieno[3.2c]pyridin-2-yl)-methyl]-L-prolinamide,hydrochloride (N—(HOOCCH₂)-D-Cha-Pro-Atp. HCl)

[0590] (step 1; coupling of the fist amino acid to Kaiser oxim resin).N-Boc-Pro-OH, (1.7 g, 8.0 mmol) and N,N′-dicyclohexylcarbodiimide (0.83g, 4.0 mmol) in dichloromethane (20 mL) were stirred for 1 hour at 0deg., the precipitate was removed and the solution was added to Kaiseroxim resin (1.0 g, 1.0 mmol, swollen in dichloromethane for 2 hoursprior to the addition of the activated proline derivative). The resinwas shaken for 16 h, the solvent was removed, the resin was washed withdichoromethane and diethyl ether (3 times each) and air dried.

[0591] (step 2; deprotection). 200mg of the N-Boc-Pro-coupled resin wasswollen for 2 hours in dichloromethane (2 ml). 4 mL of 25%trifluoroacetic acid in dichloromethane was added and shaken for 2 hoursat room temperature. The solvent was removed, the resin was washed withdichloromethane, methanol, and diethylether, and air dried.

[0592] (step 3; coupling to the second amino acid) 134 mg of the resinfrom step 2 was swollen in N,N-dimethylformamide for 30 min. The solventwas removed, and a solution of 142 mgN-Boc-N-t-butyloxycarbonylmethyl)-D-Cha-OH, 118 mg TBTU and 0,04 mLN-methyl morpholine were added to the resin. The mixture was shaken for16 h, the solvent was removed and the resin was washed withN,N-dimethylformamide, methanol, and diethylether.

[0593] (step 4, cleavage of the product from the resin using an amine).150 mg (0.1 mmol) of the resin from step 3 was swollen indichloromethane for 2 h. The solvent was removed, a mixture of4-amino-2-(aminomethyl)thieno[3,2c]pyridine hydrochloride (73c.) (67 mg,0.3 mmol) and N,N-diisopropylethylamine (0.3 mL) in dichloromethane (2-7mL) was added to the resin. The mixture was shaken for 16 hours thesolvent was removed and collected, the resin was washed withdichloromethane (1 mL). The solutions were combined, the solvent wasremoved i.vac., the residue was dissolved in ethyl acetate and extractedwith phosphated buffer (pH=7.0). The organic phase was collected, dried(magnesium sulfate), filtered, and the solvent was removed to giveN-Boc-N-(t-butyloxycarbonylmethyl)-D-Cha-Pro-Atp (11 mg, 17%), FAB-MS,m/e=643 [M+].

[0594] (step 5; deprotection) The product from step 4 was dissolved in0.5 mL 4 M HCl in dioxane and kept at 5 deg for 16 h. The solvent wasremoved i.vac., and the product triturated with diethylether to give 5mg (62%) of the target compound. Mp 170 deg (dec); FAB-MS (m/e) 487[M+].

Example 813-[4-[[3-(4-Aminothieno[3,2c]pyridin-2-yl)acryloyl](methyl)amino]piperidin-1-yl]-3-oxopropionicacid methyl ester

[0595] 81a. 3-[4-(Benzoylamino)thieno[3,2c]pyridin-2-yl]acrylic acid

[0596] A mixture of N-(2-formylthieno[3,2c]pyridin-4-yl)benzamide (3d:5.2 g, 18.5 mmol), malonic acid (3.83 g, 36.8 mmol), and piperidine(0.73 mL, 7.4 mmol) in 57 mL of pyridine was heated at 105° C. during5.5 hours. After evaporation of pyridine, water was added and 0.1 Nhydrochloric acid until pH=3. The precipitate formed was collected byfiltration, washed with water and acetone to yield a yellow compound(3.9 g).

[0597] 81b. 3-(4-Aminothieno[3,2c]pyridin-2-yl)acrylic acid

[0598] A solution of 3-[4-(benzoylamino)thieno[3,2c]pyridin-2-yl]acrylicacid (3.9 g, 12 mmol) in 4 N hydrochloric acid (760 mL) and acetic acid(240 mL) was heated at reflux during 20 hours. The reaction mixture wascooled, then the precipitate formed was filtered off and washed withdiethyl ether, and dried under vacuum to yield the title compound (2.3g).

[0599] 81c. 3Oxo-3-(4-oxopiperidin-1-yl)propionic acid methyl ester

[0600] To a solution of 4-piperidone monohydrate, hydrochloride (30.7 g;0.2 mol), sodium carbonate (42 g; 0.4 mol) in water (160 mL) was addeddichloromethane (500 mL). The mixture was stirred at room temperatureand a solution of methyl malonylchloride (26 ml; 1.2 eq) indichloromethane (100 mL) was added dropwise at room temperature, keepingthe temperature of the reaction mixture below 30° C. The pH wasmaintained at 8 at the end of the addition. The mixture was stirred 4hours at room temperature. The residue was extracted twice withdichloromethane, dried over magnesium sulfate and evaporate to yield thetitle compound (37.4 g).

[0601] 81d. 3-(4-(Methylamino)piperidin-1-yl)-3-oxopropionic acid methylester hydrochloride

[0602] To a solution of methylamine hydrochloride (67.5 g, 10 eq) inmethanol (520 mL) was added a solution of3-oxo-3-(4-oxopiperidin-1-yl)propionic acid methyl ester (20 g; 0.1 mol)in methanol (170 mL) and stepwise sodium cyanoborohydride (powder; 5.03g, 0.08 mol). The reaction mixture was stirred 24 hours at roomtemperature. After addition of a saturated water solution of sodiumcarbonate, the residue was evaporated and extracted withdichloromethane. The organic phase was dried and evaporated to yield thecrude 3-(4-(methylamino)piperidin-1-yl)-3-oxopropionic acid methyl ester(11.6 g) which was converted to the hydrochloride with hydrogen chloridein ethyl acetate/methanol.

[0603] 81e.3-[4-[[3-(4-Aminothieno[3,2c]pyridin-2-yl)acryloyl](methyl)amino]piperidin-1-yl]-3-oxopropionicacid methyl ester

[0604] To a solution of 3-(4-aminothieno[3,2c]pyridin-2-yl)acrylic acid(1 g, 3.9 mmol) in 110 mL of N,N-dimethylformamide, under nitrogen, wasadded N-ethylmorpholine (1.5 mL, 11.7 mmol), hydroxybenzotriazole (0.53g, 3.9 mmol) followed by3-(4-(methylamino)piperidin-1-yl)-3-oxopropionic acid methyl esterhydrochloride (0.98. g, 3.9 mmol) and N′-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (EDC). The reaction mixture was stirred atroom temperature during 4 hours. After evaporation ofN,N-dimethylformamide under reduced pressure, the residue was dissolvedin 200 mL of dichloromethane. The organic phase was washed water,saturated sodium hydrogencarbonate and water, dried with magnesiumsulfate, and evaporated to yield the pure the title compound afterseveral washing with diethyl ether, and drying under vacuum (600 mg).m.p.=130° C. Calculated % C: 57.58 H: 5.81 N: 13.44 Found % C: 55.78 H:5.84 N: 13.05

Example 823-[4-[[3-(4-Aminothieno[3,2c]pyridin-2-yl)acryloyl](methyl)amino]piperidin-1-yl3-oxopropionicacid dihydrochloride

[0605] A solution of3-[4-[[3-(4-aminothieno[3,2c]pyridin-2-yl)acryloyl](methyl)amino]piperidin-1-yl]-3-oxopropionicacid methyl ester (0.1 g, 2.4 mmol) in 3N hydrochloric acid (10 mL) wasstirred during 24 hours at room temperature. Hydrochloric phase wasevaporated under pressure. The residue was triturated with acetone, toyield the title compound as crystals, which were dried under vacuum withP₂O₅. (80 mg). m.p.=200° C. Calculated % C: 48.00 H: 5.09 N: 11.79 Found% C: 47.73 H: 5.12 N: 11.59

Example 833-(4-{[3-(1-amino-6-isoquinolinyl)-acryloyl](methyl)amino]piperidin-1-yl)-3-oxo-propionicacid methyl ester

[0606] Using the method described in example 81aN-(6-formyl-1-isoquinolinyl)benzamide (1f) was transformed into3-(1-(benzylamino)-6-isoquinolinyl)acrylic acid. This compound was usedto prepare 3-(1-amino-6-isoquinolinyl)acrylic acid according to theprocedure described in example 81b. Reaction of3-(1-amino-6-isoquinolinyl)acrylic acid with3-(4-(methylamino)piperidin-1-yl)-3-oxopropionic acid methyl esterhydrochloride according to the procedures described in example 81eyielded the title compound. NMR (CDCl₃) δ:1.8 (4H, m), 2.7 (1H, m), 3.0(3H, s), 3.2 (1H, m), 3.5 (2H, d), 3.7 (3H, s), 3.9 (1H, br d), 4.8 (2H,m), 4.9 (2H, br s), 7.0 (1H, d), 7.5 (1H, d), 7.65 (1H, d), 7.8 (3H, m),8.0 (1H, d).

Example 843-(4-{[3-(1-amino-6-isoquinolinyl)acryloyl]methyl)amino]piperidin-1-yl}-3-oxo-propionicacid hydrochloride

[0607] The method is the same as in example 82 but starting from3-(4-{[3-(1-amino-6-isoquinolinyl)-acryloyl](methyl)amino]piperidin-1-yl)-3-oxo-propionicacid methyl ester (Example 83) NMR (DMSO) δ:1.8 (4 H, m), 2.7-3.4 (5H,m+2s), 3.5 (2H, s), 3.9 (1H, br d), 4.3-4.9 (2H, m +br d), 7.1-8.9 (7H,m), 9.4 (2H, br s), 13.4 (1H, br s)

Example 85 (7-Methoxynaphthalen-2-yl)sulfonic acid{1-[1-amino-7-isoquinolinylmethyl]-2-oxopyrrolidin-3-(S)-yl}amidehydrochloride (85j.)

[0608] 85a. 7-Methoxy-isoquinoline

[0609] Aminoacetaldehyde dimethyl acetal (81.9 mL) was added to asolution of 60.8 mL of 3-methoxy-benzaldehyde in 600 mL of toluene. Themixture was refluxed for 5 h using a Dean-Stark trap and subsequentlycooled to 5° C. In a nitrogene atmosphere trifluoroacetic acid anhydride(209 mL) and borontrifluoride etherate (185 mL) were added in successionat such a rate, that the internal temperature was kept below 10° C.After stirring for 3 days at room temperature the the reaction mixturewas poured on ice, 250 mL 2 N hydrochloric acid added and the organiclayer extracted with 1 N hydrochloric acid. The pH value of the combinedaqueous extracts was adjusted to pH 9 by adding concentrated aqueousammonia. Extraction with ethyl acetate, followed by drying and removalof the solvent in vacuo gave 66.9 g (84%) of the title compound as alight brownish oil. EI-MS: 159 (M⁺).

[0610] 85b. 7-Methoxy-isoquinoline-N-oxide hydrochloride:

[0611] At room temperature 58 g of m-chloroperoxybenzoic acid (purity75%) were added in portions to a stirred solution of7-methoxy-isoquinoline (35.9 g) in 500 mL of dichloromethane. Stirringwas continued for 3 hours and subsequently methanol (400 mL) was added.The bulk was reduced to 300 mL and 325 mL of a saturated solution ofhydrogen chloride in diethyl ether were added. Dilution with 600 mL ofdiethyl ether afforded precipitation of yellow crystals, which wereseparated by filtration, washed with chilled diethyl ether and dried invacuo. Yield: 41.3 g (87%); m.p. 185-187° C.; (+)−FAB-MS: 176 (MH⁺−HCl).

[0612] 85c. 1-Chloro-7-methoxy-isoquinoline:

[0613] 7-Methoxy-isoquinoline-N-oxide hydrochloride (38.2 g) was addedin portions to phosphoryl chloride (275 mL) and the mixture heated at90° C. for 6 h. Excess of phosphoryl chloride was removed in vacuo. Theremaining white solid was washed with water, filtered and dried invacuo. Yield: 28.3 g (81%), m.p. 77-78° C.

[0614] 85d. 7-Methoxy-isoquinolin-1-ylamine

[0615] Liquid ammonia (220 mL) was added to a solution of 32.8 g of1-chloro-7-methoxy-isoquinoline in 420 mL of ethanol in a steel vessel.Nitrogen was pressed upon until an initial pressure of 20 atm. wasobtained. This reaction mixture was heated for 2 days at 170° C. Thesolvent was removed in vacuo and the residue dissolved in water. The pHvalue was adjusted to pH 10 by adding aqueous sodium carbonate solution,followed by extraction with ethyl acetate. The organic extract waswashed with brine and dried (Na₂SO₄). Evaporation of the solvent invacuo gave pure 7-methoxy-isoquinolin-1-ylamine as a white solid. Yield24.0 g (81%); m.p. 128-130° C.

[0616] 85e. 1-Amino-isoquinolin-7-ol

[0617] Boron tribromide (35 mL) in 50 mL of dichloromethane was addeddropwise to a stirred solution of 7-methoxy-isoquinolin-1-ylamine (21.6g) in 70 mL of dichloromethane at 10° C. After stirring for 4 d atambient temperature the reaction mixture was poured on ice and the pHadjusted to pH 9 by adding concentrated aqueous ammonia. Theprecipitated material was collected by filtration and dried in vacuo togive 19.3 g (97%) of the title compound as a light brownish solid. M.p.260° C. (decomp.); EI-MS: 160 (M⁺).

[0618] 85f. Trifluoro-methanesulfonic acid 1-amino-isoquinolin-7-ylester

[0619] A mixture of 20.0 g of I-amino-isoquinolin-7-ol and 67.2 g ofN-phenyl-bis(trifluoromethane-sulfonimid) in 300 mL of dichloromethaneand 300 mL of dioxane was cooled in an ice bath and 26.9 mL ofN,N-diisopropylethylamine added dropwise. The resulting mixture washeated for 24 h at 70° C., after which the volatiles were removed invacuo. The remaining residue was dissolved in ethyl acetate, washed withsuccessive portions of 2N NaOH, water and brine and dried (Na₂SO₄).Filtration and concentration afforded a colourless oil, which waspurified by silica chromatography (ethyl acetate/isohexane=3/7, 4/6,5/5, 6/4, 7/3) yielding 35.4 g (92%) of the title compound as yellowcrystals. M.p. 115-118° C.; EI-MS: 292 (M⁺).

[0620] 85g. 1-Amino-isoquinoline-7-carbonitrile

[0621] Palladium acetate (0.9 g) was added to a heated mixture oftrifluoromethanesulfonic acid 1-amino-isoquinolin-7-yl ester (5.8 g),zinc cyanide (2.3 g) and triphenylphosphine (1.0 g) in 75 mL ofN-methyl-pyrrolidone at 190° C. (exothermic!). Stirring was continued at190° C. for 2 h. Precipitated material was removed by filtration anddiscarded. Ethyl acetate was added and the organic mixture washed with2N aqueous ammonia, water and brine and dried (Na₂SO₄). Filtration andconcentration afforded a brownish oil, which was purified by silicachromatography (ethyl acetate/isohexane=3/7, 4/6, 5/5, 6/4, 7/3, 8/2) togive 3.2 g (94%) of the title compound as yellow crystals. M.p. 183-186°C.

[0622] 85h. 1-amino-7-(aminomethyl)isoquinoline

[0623] Liquid ammonia (210 mL) was added to a mixture of 4.2 g of1-amino-isoquinoline-7-carbonitrile and 4.0 g of Raney-Ni in 210 mL ofmethanol in a steel vessel. Hydrogen was pressed upon until an initialpressure of 100 atm. was obtained. This mixture was reacted for 16 hoursat ambient temperature, the catalyst subsequently removed by filtrationand the solvent pumped off. The residue was purified by silicachromatography (methanol/ammonia saturated methanol=85:15, 8:2) yielding1.1 g (25%) of the title compound as a light yellow solid. M.p. 114-117°C.

[0624] 85i.[1-(1-amino-7-isoquinolinylmethyl)-2-oxopyrrolidin-3-(S)yl]carbamic acidtert-butyl ester

[0625] To a solution of 0.3 g of Boc-L-Asp(H)-OBn (WO 96/40679)dissolved in 2 mL methanol was added 176 mg of1-amino-7-(aminomethyl)isoquinoline. After stirring for 30 minutes, asolution of 80 mg sodium cyanoborohydride and 90 mg zinc chloride in 2mL methanol was added. The mixture was stirred for an additional twohours. After this time, 7 mL of an aqueous 0.2 N sodium hydroxidesolution was added and the resulting mixture was concentrated. Theresidue was treated with 100 mL dichloromethane and 10 mL water,filtered and an additional 100 mL water was added to the filtrate. Theorganic layer was separated and washed with aqueous 5% sodiumhydrogencarbonate and brine. All aqueous layers were washed twice withdichloromethane. The combined organic layers were dried over sodiumsulfate and concentrated. The residue was purified by columnchromatography on silica gel eluting with a gradient of ethylacetate/dichloromethane/methanol=10/10/1 todichloromethane/methanol=10/1 to give 146 mg of the title compound.

[0626] Rf=0.3 in ethyl acetate/dichloromethane/methanol=10/10/2 (v/v) onsilica.

[0627] 85j. (7-Methoxynaphthalen-2-yl)sulfonic acid{1-[1-amino-7-isoquinolinylmethyl]-2-oxopyrrolidin-3 -(S)-yl}amidehydrochloride

[0628] To a stirred solution of 146 mg of[1-(1-amino-7-isoquinolinylmethyl)-2-oxopyrrolidin-3-(S)yl]carbamic acidtert-butyl ester in 2 mL dichloromethane and 0.1 mL methanol was added 5mL of a 3M hydrogenchloride solution in dioxane. After stirring for 50minutes at room temperature the mixture was concentrated. To the residuewas added 10 mL dichloromethane, 0.285 mL N,N-diisopropylethylamine and118 mg 7-methoxynaphthalen-2-ylsulfonyichlorideand stirred at roomtemperature for 16 hours. Dichloromethane was added and the mixture wasextracted with aqueous 5% sodium hydrogencarbonate and brine. Bothaqueous extracts were washed with dichloromethane and the combineddichloromethane extracts were dried over sodium sulfate andconcentrated. The residue was purified by column chromatography onsilica gel eluting with ethyl acetate/dichloromethane/methanol=10/10/2to give the free base. This free base was dissolved in tert-butanol, oneequivalent hydrochloric acid was added and lyophilisation gave 90 mg ofthe title compound. MS ESI+:477 (M+H).

Example 86 (7-Methoxynaphthalen-2-yl)sulfonic acid{1-[(4-amino-furo[3,2]-pyridin-2-yl)methyl]-2-oxopyrrolidin-3-(S)-yl}amidehydrochloride (86f.)

[0629] 86a. Furo[3,2-c]pyridin-4-ylamine

[0630] The procedure described to prepare 7-methoxyisoquinolin-1-ylaminewas used to convert 15.3 g of 4-chloro-furo[3,2-c]pyridine into 12.2 gof the title compound as a brownish solid. m.p. 122-124° C.; EI-MS: 134(M⁺).

[0631] 86b. 2-Iodo-furo[3,2-c]pyridin-4-ylamine

[0632] Furo[3,2-c]pyridin-4-ylamine (7.8 g) was dissolved in 175 mL ofglacial acetic acid and sodium acetate (14.3 g) added in portions(exothermic!). Subsequently a solution of iodine (44.2 g) in 150 mL oftetrahydrofuran was added dropwise and the resulting mixture allowed tostir for 4 d at ambient temperature. The mixture was poured on ice andthe pH value adjusted to pH 10 by adding 10 N sodium hydroxide solution,followed by extraction with ethyl acetate. The organic extract waswashed several times with sodium thiosulfate solution and brine, dried(Na₂SO₄) and concentrated. The crude product was purified by silicachromatography (ethyl acetate/isohexane =5/5, 7.5/2.5, 10/0) to give10.3 g (68%) of the title compound as a brownish solid. M.p. 170-173°C.; EI-MS: 260 (M⁺).

[0633] 86c. 4-Amino-furo[3,2-c]pyridine-2-carbonitrile

[0634] This compound was prepared from2-iodo-furo[3,2-c]pyridin-4-ylamine (5.2 g), zinc cyanide (3.6 g),triphenylphosphine (2.1 g) and palladium acetate (1.8 g) in 75 mL ofN-methyl-pyrrolidone using the procedure described for1-amino-isoquinoline-7-carbonitrile. Yield: 2.0 g (64%); yellow solid;m.p. 280° C. (decomp.).

[0635] 86d. 4-Amino-2-(aminomethyl)furo[3,2-c]pyridine

[0636] This compound was prepared from4-amino-furo[3,2-c]pyridine-2-carbonitrile (5.7 g), using the proceduredescribed for 1-amino-7-(aminomethyl)isoquinoline. Yield: 3.5 g (60%);yellow crystals; m.p. 142-144° C.; EI-MS: 163 (M⁺).

[0637] 86e.[1-((4-amino-furo[3,2-c]pyridin-2-yl)methylamino)-2-oxopyrrolidin-3-(S)yl]carbamicacid tert-butyl ester

[0638] Using the procedure described in example 85i 166 mg of4-amino-2-(aminomethyl)furo[3,2-c]pyridine was transformed into 150 mgof the title compound. MS ESI+:347 (M+H).

[0639] 86f. (7-Methoxynaphthalen-2-yl)sulfonic acid{1-[(4-amino-furo[3,2-c]pyridin-2-yl)methyl]-2-oxopyrrolidin-3-(S)-yl}amidehydrochloride

[0640] Using the procedure described in example 85j 150 mg of[1-((4-amino-furo[3,2-c]pyridin-2-yl)methylamino)-2-oxopyrrolidin-3-(S)yl]carbamicacid tert-butyl ester was transformed into 110 mg of the title compound.MS ESI+: 467 (M+H).

Example 87 (7-Methoxynaphthalen-2-yl)sulfonic acid{1-[1-amino-6-isoquinolinylmethyl]-2-oxopyrrolidin-3-(S)-yl}amidehydrochloride

[0641] Using the procedure described in example 85 176 mg of1-amino-6-(aminomethyl)isoquinoline was transformed into 81 mg of thetitle compound. MS ESI+: 477 (M+H).

Example 88 (7-Methoxynaphthalen-2-yl)sulfonic acid{1-[(4-amino-thieno[3,2-c]pyridin-2-yl)methyl]-2-oxopyrrolidin-3-(S)-yl}amidehydrochloride

[0642] Using the procedure described in example 85 184 mg of4-amino-2-(aminomethyl)thieno[3,2-c]pyridine was transformed into 222 mgof the title compound. MS ESI+: 483 (M+H).

Example 891-Amino-6-[(2-(3-(cyclohexylmethylsulfonylamino)-6-methyl-2-oxo-1,2-dihydropyridinyl)-1-oxo-ethyl)aminomethyl]isoquinolinehydrochloride (89b.)

[0643] 89a.[3-(cyclohexylmethylsulfonylamino)-6-methyl-2-oxo-1,2-dihydropyridinyl]-aceticacid

[0644] To a stirred solution of 0.71 g of tert-butyl[3-amino-6-methyl-2-oxo-1,2-dihydropyridinyl]-acetate (WO 97/01338) at0° C. under a nitrogen atmosphere was added 0.60 mLcyclohexyl-methanesulfonyl chloride (J. F. King et al J. Am. Chem. Soc.114, 1743 (1992)). After stirring at room temperature for 3 hours thereaction mixture-was concentrated. The residue was dissolved in ethylacetate, washed successively with 0.1 N hydrochloric acid, water andbrine, dried over sodium sulphate and concentrated. The residue wasdissolved in a mixture of 0.2 mL water and 1.8 mL trifluoroacetic acid.After stirring at room temperature for 4 h, water and ethyl acetate wereadded. The organic layer was separated, washed twice with 0.1 Nhydrochloric acid, dried over sodium sulphate and concentrated. Thesolid residue was washed with a cold dichloromethane/diethyl ethermixture to give 0.72 g of the title compound.

[0645] TLC: Rf=0.25, dichloromethane/methanol=9/1 v/v on silica.

[0646] 89b.1-Amino-6-[(2-(3-(cyclohexylmethylsulfonylamino)-6-methyl-2-oxo-1,2-dihydropyridinyl)-1-oxo-ethyl)aminomethyl]isocuinolinehydrochloride

[0647] To a stirred solution of 0.14 g of[3-(cyclohexylmethylsulfonylamino)-6-methyl-2-oxo-1,2-dihydropyridinyl]-aceticacid and 0.07 g of 1-amino-6-(aminomethyl)isoquinoline in 5 mLdichloromethane and 2 mL N,N-dimethylformamide was added 0.19 g2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate(TBTU). After 16 h at room temperature additional 50 mg TBTU and 0.10 mLN-methyimorpholine were added and the mixture was stirred at roomtemperature for an additional 16 h. Dichloromethane was added and thereaction mixture was washed with aqueous saturated sodiumhydrogencarbonate and water. Both aqueous washes were extracted threetimes with dichloromethane. The combined organic phases were dried oversodium sulphate and concentrated. The residue was purified bychromatography on silica gel (eluent: dichloromethane/methanol=9/1 v/v)to yield the free base. This free base was dissolved in a mixture oft-butanol/water=1/1 (v/v) and one equivalent hydrochloric acid wasadded. Lyophilisation yielded 135 mg of the title compound. MS ESI+:498(M+H).

Example 901-Amino-6-[(2-(3-(benzylsulfonylamino)-6-methyl-2-oxo-1,2-dihydropyridinyl)-1-oxo-ethyl)aminomethyl]isoquinolinehydrochloride (89b.)

[0648] The procedure decribed in example 89 was used to convert 168 mgof [3-(benzylsulfonylamino)-6-methyl-2-oxo-1,2-dihydropyridinyl]-aceticacid (WO 97/01338) into 99 mg of the title compound. MS ESI+492 (M+H).

Example 91(2-{2-(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-oxo-1-(R)-phenyl-ethyl)-carbamicacid isopropyl ester hydroacetate (91d)

[0649] 91a. (R)-Isopropoxy-carbonylamino-phenyl-acetic acid

[0650] A mixture of(D)-phenylglycine (5.0 g) in 66 mL of 1 N NaOH wascooled in an ice bath, 33 mL of isopropyl chloroformate (1 M solution intoluene) added dropwise and the resulting mixture was stirred for 16 hat ambient temperature. After adding aqueous NaOH (pH 11) the organiclayer was discarded. KHSO₄ was added and the acidified aqueous solutionextracted with ethyl acetate several times. Evaporation of the solventin vacuo gave pure (R)-isopropoxy-carbonylamino-phenyl-acetic acid.Yield: 6.3 g (81%); white solid; m.p. 123-127° C.; (−)-APCI-MS: 236([M−H]⁻).

[0651] 91b.1-[(R)-Isopropoxycarbonylamino-phenyl-acetyl]-pyrrolidine-2-(S)-carboxylicacid methyl ester

[0652] A mixture of (R)-Isopropoxycarbonylamino-phenyl-acetic acid (0.74g), (L)-proline methyl ester hydrochloride (0.51 g), N-methyl morpholine(1.00 mL) and 1.00 g of2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate(TBTU) in 10 mL of dry N,N-dimethyl formamide was stirred for 3 h atambient temperature. The volatiles were pumped off and water was addedfollowed by extraction with ethyl acetate and concentration of theorganic layer. The remaining residue was purified by silicachromatography (ethyl acetate/isohexane=3/1) to give 95 g (91%) of thetitle compound as a colorless oil. (+)-APCI-MS: 349 (MH⁺).

[0653] 91c.1-[(R)-Isopropoxycarbonylamino-phenyl-acetyl]-pyrrolidine-2-(S)-carboxylicacid

[0654] A mixture of1-[(R)-isopropoxycarbonylamino-phenyl-acetyl]-pyrrolidine-2-(S)-carboxylicacid methyl ester (0.95 g) and 0.13 g of LiOH in 20 mL of methanol and 5mL of water was stirred for 16 h at ambient temperature. KHSO₄ wasadded, the volatiles were pumped off followed by addition of water,extraction with ethyl acetate and concentration of the organic layer togive 0.76 g (83%) of the title compound as a white solid. (+)-APCI-MS:335 (MH⁺).

[0655] 91d.(2-{2-(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-oxo-1-(R)-phenyl-ethyl)-carbamic acid isopropyl ester hydroacetate

[0656] A mixture of1-[(R)-isopropoxycarbonylamino-phenyl-acetyl]-pyrrolidine-2-(S)-carboxylicacid (0.20 g), 6-aminomethyl-isoquinolin-1-ylamine (0.6 mmol), N-methylmorpholine (0.2 mL) and 0.19 g of2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate(TBTU) in 3 mL of dry N,N-dimethyl formamide was stirred for 3 h atambient temperature. 0.120 mL CH₃COOH were added, the mixture wasconcentrated and the remaining residue was purified by HPLC (RP-18;H₂O/CH₃OH 95/5→0/100) to give 0.10 g (35%) of the title compound as acolorless oil. (+)-APCI-MS: 490 (MH⁺).

Example 92(1-(R)-{2-(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-carbonyl}-2-isopropyl-sulfanyl-2-methyl-propyl)-carbamicacid ethyl ester hydroacetate (92e)

[0657] 92a. 2-(R)-Amino-3-isopropylsulfanyl-3-methyl-butyric acid

[0658] NaH (1.3 g) was added in portions to 70 mL ethanol while cooling.Subsequently D-penicillamine (4.0 g) was added. The mixture keptstirring for 5 minutes at 5° C., after which isopropyl iodide (2.8 mL)was added. The mixture was allowed to stir 16 h at ambient temperature.2 N HCl was added followed by evaporation of the volatiles to give 11.9g crude 2-(R)-amino-3-isopropylsulfanyl-3-methyl-butyric acid, which wasused in the next step without further purification

[0659] 92b.2-(R)-Ethoxycarbonylamino-3-isopropylsulfanyl-3-methyl-butyric acid

[0660] 2-(R)-Amino-3-isopropylsulfanyl-3-methyl-butyric acid (approx. 27mmol; crude material from the previous step) was dissolved in 40 mL ofwater and 40 mL of dioxane and pH 9.5 was adjusted by adding aqueousNaOH. Ethyl chloroformate (3.3 mL) was slowly added at 5° C. whilemaintaining pH 9.5 by addition of the appropriate amount of aqueousNaOH. Stirring was continued for 6 h at room temperature. Aqueous NaOHwas added (pH 11) and the dioxane removed in vacuo followed byextraction of the remaining aqueous solution with ethyl acetate. KHSO₄was added and the acidified aqueous solution extracted with ethylacetate several times. Evaporation of the solvent in vacuo gave a yellowoil, which was purified by HPLC (RP-18; pH 2.3; H₂O/CH₃OH 3/7) to give1.50 g (21%) of the title compound as a colorless oil. (+)-APCI-MS: 264(MH⁺).

[0661] 92c.1-[2-(R)-Ethoxycarbonylamino-3-isopropylsulfanyl-3-methyl-butyryl-]-pyrrolidine-2-(S)-carboxylicacid methyl ester

[0662] A mixture of (L)-proline methyl ester hydrochloride (0.28 g),2-(R)-ethoxycarbonylamino-3-isopropylsulfanyl-3-methyl-butyric acid(0.44 g), N-methyl morpholine (0.73 mL) and 0.54 g of2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate(TBTU) in 10 mL of dry N,N-dimethylformamide was stirred for 16 h atambient temperature. The volatiles were pumped off and water was addedfollowed by extraction with ethyl acetate and concentration of theorganic layer. The remaining residue was purified by silicachromatography (ethyl acetate/isohexane=1/1) to give 0.45 g (72%) of thetitle compound as a colorless oil. (+)-APCI-MS: 375 (MH⁺).

[0663] 92d.1-[2-(R)-Ethoxycarbonylamino-3-isopropylsulfanyl-3-methyl-butyryl]-pyrrolidine-2-(S)-carboxylicacid

[0664] A mixture of1-[2-(R)-ethoxycarbonylamino-3-isopropylsulfanyl-3-methyl-butyryl-]-pyrrolidine-2-(S)-carboxylicacid methyl ester (0.45 g) and 0.06 g of LiOH in 6 mL of methanol and 1mL of water was stirred for 16 h at ambient temperature. KHSO₄ wasadded, the volatiles were pumped off followed by addition of water,extraction with ethyl acetate and concentration of the organic layer togive 0.34 g (79%) of the title compound as a colorless oil. (+)-APCI-MS:361 (MH⁺).

[0665] 92e.(1-(R)-{2-(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-carbonyl}-2-iso-propylsulfanyl-2-methyl-propyl)-carbamicacid ethyl ester hydroacetate

[0666] A mixture of1-[2-(R)-ethoxycarbonylamino-3-isopropylsulfanyl-3-methyl-butyryl]-pyrrolidine-2-(S)-carboxylicacid (0.160 g), 6-aminomethyl-isoquinolin-1-ylamine (0.078 g), N-methylmorpholine (0.14 mL) and 0.14 g of2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate(TBTU) in 3 mL of dry N,N-dimethyl formamide was stirred for 3 h atambient temperature. 1.000 mL CH₃COOH were added, the mixture wasconcentrated and the remaining residue was purified by HPLC (RP-18;H₂O/CH₃OH 95/5→0/100) to give 0.17 g (66%) of the title compound as acolorless oil. (+)-APCI-MS: 516 (MH⁺).

Example 93(2-{2-(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-oxo-1-(R)-phenyl-ethyl)-carbamicacid isobutylester hydroacetate (93d)

[0667] 93a. (R)-Isobutoxy-carbonylamino-phenyl-acetic acid

[0668] The procedure described for example 91a was used. Reaction of 2.0g of (D)-phenylglycine and 2.0 mL of isobutyl chloroformate gave 0.4 g(12%) of the title compound as a colorless oil. (−)-APCI-MS: 250([M−H]⁻).

[0669] 93b.1-[(R)-Isobutoxycarbonylamino-phenyl-acetyl]-pyrrolidine-2-(S)-carboxylicacid methyl ester

[0670] The procedure described for example 91b was used. From 0.40 g of(R)-isobutoxycarbonylamino-phenyl-acetic acid and 0.26 g of (L)-prolinemethyl ester hydrochloride 0.45 g (78%) of the title compound wasobtained as a colorless oil (+)-APCI-MS: 363 (MH⁺).

[0671] 93c.1-((R)-Isobutoxycarbonylamino-phenyl-acetyl]-pyrrolidine-2-(S)-carboxylicacid

[0672] Starting with 0.45 g of1-[(R)-isobutoxycarbonylamino-phenyl-acetyl]-pyrrolidine-2-(S)-carboxylicacid methyl ester using the procedure described for example 91c gave0.38 g (88%) of the title compound as a white solid. (+)-APCI-MS: 349(MH⁺).

[0673] 93d.(2-{2-(S)-](1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-oxo-1-(R)-phenyl-ethyl)-carbamicacid isobutylester hydroacetate

[0674] The procedure described in example 91d was used to transform0.100 g of1-[(R)-isobutoxycarbonylamino-phenyl-acetyl]-pyrrolidine-2-(S)-carboxylicacid into 0.045 g (32%) of the title compound. (+)-APCI-MS: 504 (MH⁺).

Example 94(2-{2-(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-oxo-1-(R)-phenyl-ethyl)-carbamicacid ethyl ester hydroacetate (94d)

[0675] 94a. (R)-Ethoxy-carbonylamino-phenyl-acetic acid

[0676] The procedure described for example 91a was used. 2.0 g of(D)-phenylglycine as starting material gave 2.1 g (71%) of the titlecompound as a white solid. M.p. 143-150° C.; (−)-APCI-MS: 222 ([M−H]⁻).

[0677] 94b.1-[(R)-Ethoxycarbonylamino-phenyl-acetyl]-pyrrolidine-2-(S)-carboxylicacid methyl ester

[0678] Reaction of 2.0 g of (R)-ethoxycarbonylamino-phenyl-acetic acidand 1.50 g of (L)-proline methyl ester hydrochloride according to theprocedure described for example 91b gave 1.70 g (57%) of the titlecompound as a colorless oil. (+)-APCI-MS: 335 (MH⁺).

[0679] 94c.1-[(R)-Ethoxycarbonylamino-phenyl-acetyl]-pyrrolidine-2-(S)-carboxylicacid

[0680] Saponification of 1.70 g of1-[(R)-ethoxycarbonylamino-phenyl-acetyl]-pyrrolidine-2-(S)-carboxylicacid methyl ester according to the procedure described in example 91cgave 1.44 g (88%) of the title compound as a white solid. (m.p. 163-166°C.). (+)-APCI-MS: 321 (MH⁺).

[0681] 94d.(2-}2-(S)-(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-oxo-1-(R)-phenyl-ethyl)-carbamicacid ethyl ester hydroacetate

[0682] The procedure described for example 91d was used to convert 0.35g of1-[(R)-ethoxycarbonylamino-phenyl-acetyl]-pyrrolidine-2-(S)-carboxylicacid into 0.31 g (53%) of the title compound. (+)-APCI-MS: 476 (MH⁺)

Example 95(1-(R)-{2-(S)-](1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-carbonyl}-2-isopropylsulfanyl-2-methyl-propyl)-carbamicacid isopropyl ester hydroacetate (95d)

[0683] 95a.2-(R)-Isopropoxycarbonylamino-3-isopropylsulfanyl-3-methyl-butyric acid

[0684] 2-(R)-Amino-3-isopropylsulfanyl-3-methyl-butyric acid (17 mmol;crude material obtained from D-penicillamine and isopropyl iodide) wasdissolved in 40 mL of water and 40 mL of dioxane and pH 9.5 was adjustedby adding aqueous NaOH. Isopropyl chloroformate (22 mL; 1 M solution intoluene) was slowly added at 5° C. while maintaining pH 9.5 by additionof the appropriate amount of aqueous NaOH. Stirring was continued for 6h at room temperature. Aqueous NaOH was added (pH 11) and the dioxaneremoved in vacuo followed by extraction of the remaining aqueoussolution with ethyl acetate. KHSO₄ was added and the acidified aqueoussolution extracted with ethyl acetate several times. Evaporation of thesolvent in vacuo gave a yellow oil, which was purified by HPLC (RP-18;pH 2.3; H₂O/CH₃OH 3/7) to give 3.0 g (64%) of the title compound as acolorless oil. (−)-APCI-MS: 276 ([M−H]⁻).

[0685] 95b.1-[2-(R)-Isopropoxycarbonylamino-3-isopropylsulfanyl-3-methyl-butyryl-]-pyrrolidine-2-(S)-carboxylicacid methyl ester

[0686] Reaction of 0.54 g of (L)-proline methyl ester hydrochloride and0.89 g of2-(R)-isopropoxy-carbonylamino-3-isopropylsulfanyl-3-methyl-butyric acidaccording to the procedure described for example 91b afforded 1.0 g(88%) of the title compound as a colorless oil. (+)-APCI-MS: 389 (MH⁺)

[0687] 95c.1-[2-(R)-Isopropoxycarbonylamino-3-isopropylsulfanyl-3-methyl-butyryl]-pyrrolidine-2-(S)-carboxylicacid

[0688] Saponification of 1.10 g of1-[2-(R)-isopropoxycarbonylamino-3-isopropylsulfanyl-3-methyl-butyryl-]-pyrrolidine-2-(S)-carboxylicacid methyl ester according to the procedure described in example 91cgave 0.59 g (79%) of the title compound as a white solid. M.p. 128-130°C.; (−)- APCI-MS: 373 ([M−H]⁻)

[0689] 95d.(1-(R)-{2-(S)-](1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-carbonyl}-2-iso-propylsulfanyl-2-methyl-propyl)-carbamicacid ethyl ester hydroacetate

[0690] The procedure described for example 91d was used to convert 0.13g of1-[2-(R)-isopropoxycarbonylamino-3-isopropylsulfanyl-3-methyl-butyryl]-pyrrolidine-2-(S)-carboxylicacid into 0.13 g of the title compound. (+)-APCI-MS: 530 (MH⁺)

Example 96(3-{2-(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-(R)-ethoxy-carbonylamino-1,1-dimethyl-3-oxo-propylsulfanyl)-aceticacid tert.-butyl ester hydroacetate (96d)

[0691] 96a.3-tert.-Butoxycarbonylmethylsulfanyl-2-(R)-ethoxycarbonylamino-3-methyl-butyricacid

[0692] NaH (0.81 g; 35.2 mmol) was added in portions to 40 mL ethanolwhile cooling. Subsequently D-penicillamine (2.50 g) was added. Themixture kept stirring for 5 minutes at 5° C., after which bromoaceticacid tert.-butyl ester (2.55 mL) was added. The mixture was allowed tostir 16 h at ambient temperature and subsequently hydrolyzed with 2 NHCl. The solvents were distilled off, the residue was redissolved in 50mL of water and 50 mL of dioxane and pH 9.5 was adjusted by addingaqueous NaOH. Ethyl chloroformate (2.1 mL) was slowly added at 5° C.while maintaining pH 9.5 by addition of the appropriate amount ofaqueous NaOH. Stirring was continued for 6 h at room temperature.Aqueous NaOH was added (pH 10) and the dioxane removed in vacuo followedby extraction of the remaining aqueous solution with ethyl acetate.KHSO₄ was added and the acidified aqueous solution extracted with ethylacetate several times. Evaporation of the solvent in vacuo gave acolorless oil, which was purified by HPLC (RP-18; pH 2.3, H₂O/CH₃OH 3/7)to give 0.86 g (15%) of the title compound as a colorless oil.(+)-APCI-MS: 358 (MNa⁺).

[0693] 96b. 1-[3-tert-Butoxycarbonylmethylsulfanyl-2-(R)-ethoxycarbonylamino-3-methyl-butyryl]-pyrrolidine-2-(S)-carboxylicacid methyl ester

[0694] Reaction of 0.44 g of (L)-proline methyl ester hydrochloride and0.86 g of3-tert.-butoxycarbonylmethylsulfanyl-2-(R)-ethoxycarbonylamino-3-methyl-butyricacid according to the procedure described in example 91b yielded 0.60 g(52%) of the title compound as a colorless oil (+)-APCI-MS: 474 (MH⁺).

[0695] 96c.1-[3-tert.-Butoxycarbonylmethylsulfanyl-2-(R)-ethoxycarbonylamino-3-methyl-butyryl]-pyrrolidine-2-(S)-carboxylicacid

[0696] Saponification of 0.60 g of1-[3-tert.-butoxycarbonylmethylsulfanyl-2-(R)-ethoxycarbonylamino-3-methyl-butyryl]-pyrrolidine-2-(S)-carboxylicacid methyl ester according to the procedure described in example 91cafforded 0.080 g(14%); of the title compound as a colorless oil;(+)-APCI-MS: 433 (MH⁺).

[0697] 96d.(3-{2-(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-(R)-ethoxy-carbonylamino-1.1-dimethyl-3-oxo-propylsulfanyl)-aceticacid tert.-butyl ester hydroacetate

[0698] The procedure described for example 91d. was used to convert0.080 g of1-[3-tert.-Butoxycarbonylmethylsulfanyl-2-(R)-ethoxycarbonylamino-3-methyl-butyryl]-pyrrolidine-2-(S)-carboxylicacid into 0.071 g (61%) of the title compound. (+)-APCI-MS: 588 (MH⁺).

Example 971-(2-(R)-Acetylamino-3-biphenyl-4yl-propionyl)-pyrrolidin-2-(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide (97f)

[0699] 97a. (D)-N-Acetyl-4-bromo-phenylalanine ethyl ester

[0700] (D,L)-N-Acetyl-4-bromo-phenylalanine ethyl ester (14.1 g) wasdissolved in a mixture of 200 mL dimethyl sulfoxide, 150 mL of water and40 mL of 1 M aqueous KCl (pH 4). At 39° C. a solution of subtilisine(0.13 g) in 10 mL of water was added and the reaction mixture wasallowed to stirr for one hour while maintaining pH 7 by continousaddition of the appropriate amount of 1 N NaOH. The mixture was pouredinto a mixture of 2000 mL of water and 23 mL of 1 N NaOH. Extractionwith ethyl acetate followed by evaporation of the solvent in vacuo gave5.6 g (80%) of the title compound as a white solid. M.p. 106-110° C.;(+)-APCI-MS: 315 (MH⁺).

[0701] 97b. (D)-N-Acetyl-4-phenyl-phenylalanine ethyl ester

[0702] A mixture of (D)-N-acetyl-4-bromo-phenylalanine ethyl ester (3.14g), (1.80 g) phenyl boronic acid, sodium carbonate (2.12 g), Pd(OAc)₂(0.11 g), tri-o-tolylphosphine (0.300 g) in 60 mL of dimethoxyethane and10 mL of water was heated for 3 h at 90° C. Extraction with ethyl esterfollowed by removal of the solvent in vacuo gave 2.5 g (80%) of thetitle compound as a white solid. M.p. 161-164° C.; (+)-APCI-MS: 312(MH⁺).

[0703] 97c. (D)-N-Acetyl-4-phenyl-phenylalanine

[0704] A mixture of (D)-N-acetyl-4-phenyl-phenylalanine ethyl ester(1.55 g) and 0.16 g of LiOH in 20 mL of methanol and 5 mL of water wasstirred for 2 h at ambient temperature. KHSO₄ was added, the volatileswere pumped off followed by addition of water, extraction with ethylacetate and removal of the solvent. Yield: 1.14 g (81%) of the titlecompound as a white solid. M.p. 219-223° C.; (−)-APCI-MS: 282 ([M−H]⁻).

[0705] 97d.1-(2-(R)-Acetylamino-3-biphenyl-4yl-propionyl)-pyrrolidin-2-(S)-carboxylicacid benzyl ester

[0706] Reaction of 0.41 g of (L)-proline benzyl ester and 0.57 gof(D)-N-acetyl-4-phenyl-phenylalanine according to the proceduredescribed in example 91b yielded 0.54 g (57%) of the title compound as awhite solid. (+)-APCI-MS: 471 (MH⁺).

[0707] 97e.1-(2-(R)-Acetylamino-3-biphenyl-4yl-propionyl)-pyrrolidin-2-(S)-carboxylicacid

[0708]1-(2-(R)-Acetylamino-3-biphenyl-4yl-propionyl)-pyrrolidin-2-(S)-carboxylicacid benzyl ester (0.50 g) were dissolved in 30 mL of methanol andhydrogenated for 4 h at room temperature using 0.3 g palladium oncharcoal (10%) as catalyst. Filtration followed by concentration invacuo affords 0.37 g (91%) of the title compound as a white solid.(+)-APCI-MS: 381 (MH⁺).

[0709] 97f.1-(2-(R)-Acetylamino-3-biphenyl-4yl-propionyl)-pyrrolidin-2-(S)-carboxylicAcid (1-amino-isoquinolin-6-ylmethyl)-amide

[0710] A mixture of1-(2-(R)-acetylamino-3-biphenyl-4yl-propionyl)-pyrrolidin-2-(S)-carboxylicacid (0.120 g), 6-aminomethyl-isoquinolin-1-ylamine (0.058 g), N-methylmorpholine (0.11 mL) and 0.100 g of2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate(TBTU) in 4.5 mL of dry N,N-dimethyl formamide was stirred for 3 h atambient temperature. The mixture was concentrated and the remainingresidue was purified by silica chromatography (dichloro-methane/methanol8/2) to give 0.120 g (70%) of the title compound as a light yellowsolid. (+)-APCI-MS: 536 (MH⁺).

Example 98(2-{2-(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1-(R)-(4-methoxybenzyl)-2-oxo-ethyl)-carbamicacid ethyl ester (98d).

[0711] 98a. (D)-N-Ethoxycarbonyl-4-methoxy-phenylalanine

[0712] A mixture of (D)-4-methoxyphenylalanine hydrochloride (1.16 g) in15 mL of 1 N NaOH and 10 mL of dichloromethane was cooled in an icebath, 0.48 mL of ethyl chloroformate were added dropwise and theresulting mixture was stirred for 2 h at room temperature. After addingaqueous NaOH (pH 10) the organic layer was discarded. KHSO₄ was added(pH 3) and the aqueous solution was extracted with dichloromethaneacetate. Evaporation of the solvent in vacuo gave 1.00 g (76%) of thetitle compound as a colorless oil. (−)-APCI-MS: 266 ([M−H]⁻).

[0713] 98b.1-(2-(R)-Ethoxycarbonylamino-3-(4-methoxyphenyl)-propionyl)-pyrrolidin-2-(S)-carboxylicacid benzyl ester

[0714] Reaction of 0.63 g of (L)-proline benzyl ester hydrochloride and0.70 g of (D)-N-ethoxycarbonyl-4-methoxy-phenylalanine according to theprocedure described in example 91b yielded 0.98 g (81%) of the titlecompound as a colorless oil. (+)-APCI-MS. 455 (MH⁺).

[0715] 98c.1-(2-(R)-Ethoxycarbonylamino-3-(4-methoxyphenyl)-propionyl)-pyrrolidin-2-(S)-carboxylicacid

[0716] Hydrogenation of 0.82 g of1-(2-(R)-ethoxycarbonylamino-3-(4-methoxyphenyl)-propionyl)-pyrrolidin-2-(S)-carboxylicacid benzyl ester according to the procedure described in example 97eafforded 0.61 g (93%) of the title compound as a white solid.(+)-APCI-MS: 365 (MH⁺).

[0717] 98d.(2-{2-(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1-(R)-(4-methoxybenzyl)-2-oxo-ethyl)-carbamicacid ethyl ester

[0718] Using the procedure described in example 97f 0.115 g of1-(2-(R)-ethoxycarbonylamino-3-(4-methoxyphenyl)-propionyl)-pyrrolidin-2-(S)-carboxylicacid was converted into 0.130 g (79%) of the title compound.(+)-APCI-MS: 520 (MH⁺).

Example 99(3-{2-(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-(R)-ethoxy-carbonylamino-1,1-dimethyl-3-oxo-propylsulfanyl)-aceticacid hydrotrifluoroacetate

[0719] A mixture of(3-{2-(S)-[(1-amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-(R)-ethoxycarbonylamino-1,1-dimethyl-3-oxo-propylsulfanyl)-aceticacid tert.-butyl ester hydro-acetate (0.031 g; 0.048 mmol) and 1 mLtrifluoroacetic acid in 1 mL of dichloromethane was stirred for 3 h atambient temperature. The volatiles were pumped off followed bylyophilization to give 0.016 g (52%) of the title compound as a whitesolid. (+)-APCI-MS: 532 (MH⁺).

Example 100(2-{2-(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1-(R)-cyclohexyl-2-oxo-ethyl)-carbamicacid ethyl ester hydroacetate (100d)

[0720] 100a. (R)-Ethoxy-carbonylamino-cyclohexyl-acetic acid

[0721] Using to the procedure described in example 91d, 2.85 g of(D)-cyclohexylglycine gave 1.1 g (33%) of the title compound as acolorless oil. (−)-APCI-MS: 228 ([M−H]⁻).

[0722] 100b.1-[(R)-Ethoxycarbonylamino-cyclohexyl-acetyl-pyrrolidine-2-(S)-carboxyicacid benzyl ester

[0723] Reaction of 1.1 g of (R)-ethoxycarbonylamino-cyclohexyl-aceticacid and 1.16 g of (L)-proline benzyl ester hydrochloride according tothe procedure described in example 91b gave 1.50 g (75%) of the titlecompound as a colorless oil. (+)-APCI-MS: 417 (MH⁺).

[0724] 100c.1-[(R)-Ethoxycarbonylamino-cyclohexyl-acetyl]-pyrrolidine-2-(S)-carboxylicacid

[0725] Hydrogenation of 1.5 g of1-[(R)-ethoxycarbonylamino-cyclohexyl-acetyl]-pyrrolidine-2-(S)-carboxylicacid benzyl ester according to the procedure described for example 97eafforded 1.13 g (96%) of the title compound as a white solid.(+)-APCI-MS: 327 (MH⁺).

[0726] 100d.(2-{2-(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-oxo-1-(R)-cyclohexyl-ethyl)-carbamic acid ethyl ester hydroacetate

[0727] Using the procedure described in example 91d 0.350 g of1-[(R)-ethoxycarbonylamino-cyclohexyl-acetyl]-pyrrolidine-2-(S)-carboxylicacid was converted into 0.430 g (83%) of the title compound.(+)-APCI-MS: 482 (MH⁺).

Example 1011-[2-(R)-Acetylamino-3-(4-methoxyphenyl)-propionyl)-pyrrolidin-2-(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide (101c)

[0728] 101a.1-(2-(R)-Acetylamino-3-(4-methoxyphenyl)-propionyl)-pyrrolidin-2-(S)-carboxylicacid benzyl ester

[0729] Reaction of 0.48 g of (D)-N-acetyl-4-methoxy-phenylalanine (; 2.0mmol) and 0.41 g of (L)-proline benzyl ester according to the proceduredescribed in example 91b afforded 0.35 g (41%) of the title compound asa colorless oil. (+)-APCI-MS: 425 (MH⁺).

[0730] 101b.1-(2-(R)-Acetylamino-3-(4-methoxyphenyl)-propionyl)-pyrrolidin-2-(S)-carboxylicacid

[0731] Hydrogenation 0.35 g of1-(2-(R)-acetylamino-3-(4-methoxyphenyl)-propionyl)-pyrrolidin-2-(S)-carboxylicacid benzyl ester using to the procedure described for example 97e gave0.27 g (100%) of the title compound as a colorless oil. (+)-APCI-MS: 335(MH⁺).

[0732] b 100c.1-[2-(R)-Acetylamino-3-(4-methoxyphenyl)-propionyl)-pyrrolidin-2-(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide

[0733] Using the procedure described for example 97f 0.140 g of1-(2-(R)-acetylamino-3-(4-methoxyphenyl)-propionyl)-pyrrolidin-2-(S)-carboxylicacid was converted into 0.160 g (80%) of the title compound.(+)-APCI-MS: 490 (MH⁺).

Example 102[2-{2-(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1-(R)-(4-methoxybenzyl)-2-oxo-ethylamino]-aceticacid ethyl ester hydrochloride (102h)

[0734] 102a. (D)-4-Methoxy-phenylalanine benzyl ester hydrochloride

[0735] (D)-4-Methoxy-phenylalanine hydrochloride (7.0 g),p-toluenesulphonic acid (6.9 g) and benzyl alcohol (15.0 mL) in 100 mLof toluene were refluxed for 4 h using a Dean-Stark trap, the toluenedistilled off and the residue redissolved in ethyl acetate followed byextraction with 2 N NaOH. Then HCl in diethyl ether was added and theprecipitated material collected by filtration. Yield: 7.2 g (75%); whitesolid; m.p. 207-211° C.; (+)-APCI-MS: 286 (MH⁺).

[0736] 102b.N-Ethoxycarbonylmethyl-2-(R)-amino-3-(4-methoxyphenyl)-propionic acidbenzyl ester

[0737] (D)-4-Methoxy-phenylalanine benzylester hydrochloride (1.7 g) wasdissolved in 7 mL of dry N,N-dimethyl formamide. Bromoacetic acid ethylester (1.7 mL) and triethylamine (2.6 mL) were added in succession andthe reaction mixture was allowed to stir for 16 h at room temperature.The mixture was poured on ice and extracted with ethyl acetate. Thesolvent was removed in vacuo and the residue was purified by silicachromatography (isohexane/ethyl acetate 1/1) to give 1.75 g (78%) of thetitle compound as a light yellow oil. (+)-APCI-MS: 372 (MH⁺).

[0738] 102c.N-Ethoxycarbonylmethyl-N-tert.-butoxycarbonyl-2-(R)-amino-3-(4-methoxyphenyl)-propionicacid benzyl ester

[0739] (N-Ethoxycarbonylmethyl-2-(R)-amino-3-(4-methoxyphenyl)-propionicacid benzyl ester (1.7 g) was dissolved in 7 mL of water and a solutionof BOC anhydride (1.22 g) in 7 mL of dioxane was added. The mixture wasallowed to stir for 16 h while pH 9.5 was maintained by continiousaddition of the appropriate amounts of 0.1 N NaOH. The dioxane wasdistilled off followed by extraction with ethyl acetate, removal of thesolvent in vacuo and purification of the remaining residue by silicachromatography (isohexane/ethyl acetate 1/1). Yield: 1.50 g (68%);colorless oil; (−)-APCI-MS: 470 ([M−H]⁻).

[0740] 102d.N-Ethoxycarbonylmethyl-N-tert-butoxycarbonyl-2-(R)-amino-3-(4-methoxyphenyl)-propionicacid

[0741]N-Ethoxycarbonylmethyl-N-tert.-butoxycarbonyl-2-(R)-amino-3-(4-methoxyphenyl)-propionicacid benzyl ester (1.50 g) were dissolved in 40 mL of methanol andhydrogenated for 4 h at room temperature using 0.3 g palladium oncharcoal (10%) as catalyst. Filtration followed by concentration invacuo affords 1.05 g (87%) of the title compound as a colorless oil.(−)-APCI-MS: 381 ([M−H]⁻).

[0742] 102e.{[2-{2-(S)-[Benzyloxy-carbonyl]-pyrrolidin-1-yl}-1-(R)-(4-methoxybenzyl)-2-oxo-ethyl]-tert.-butoxycarbonyl-amino}-aceticacid ethyl ester

[0743] A mixture of (L)-proline benzyl ester hydrochloride (0.49 g),N-Ethoxycarbonylmethyl-N-tert.-butoxycarbonyl-2-(R)-amino-3-(4-methoxyphenyl)-propionicacid (0.760 g), N-methyl morpholine (0.93 mL) and 0.65 g of2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium tetra-fluoroborate(TBTU) in 10 mL of dry N,N-dimethyl formamide was stirred for 16 h atambient temperature. The volatiles were pumped off and water was addedfollowed by extraction with ethyl acetate and concentration of theorganic layer. The remaining residue was purified by silicachromatography (isohexane/ethyl acetate 1/2). Yield: 0.99 g (86%);colorless oil; (+)-APCI-MS: 569(MH⁺).

[0744] 102f. {[2-{2-(S)-Carboxy-pyrrolidin-1-yl}-1-(R)-(4-methoxybenzyl)-2-oxo-ethyl]-tert.-butoxycarbonyl-amino}-aceticacid ethyl ester

[0745] 0.78 g of{[2-{2-(S)-[Benzyloxy-carbonyl]-pyrrolidin-1-yl}-1-(R)-(4-methoxybenzyl)-2-oxo-ethyl]-tert.-butoxycarbonyl-amino}-aceticacid ethyl ester were dissolved in 50 mL of methanol and hydrogenatedfor 3 h at room temperature using 0.4 g palladium on charcoal (10%) ascatalyst. Filtration followed by concentration in vacuo affords 0.58 g(89%) of the title compound as a colorless oil. (−)-APCI-MS: 477([M−H]⁻).

[0746] 102g.{[2-{2-(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1-(R)-(4-methoxybenzyl)-2-oxo-ethyl]-tert.-butoxycarbonyl-amino}-aceticacid ethyl ester

[0747] A mixture of{[2-{2-(S)-Carboxy-pyrrolidin-1-yl}-1-(R)-(4-methoxybenzyl)-2-oxo-ethyl]-tert.-butoxycarbonyl-amino}-aceticacid ethyl ester (0.240 g), 6-aminomethyl-isoquinolin-1-ylamine (0.092g), N-methyl morpholine (0.170 mL) and 0.160 g of2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate(TBTU) in 8 mL of dry N,N-dimethyl formamide was stirred for 3 h atambient temperature. Removal of the solvent in vacuo followed bypurification of the remaining residue by silica chromatography(dichloromethane/methanol 80/20) gave 0.310 g (100%) of the titlecompound as a light yellow solid. (+)-APCI-MS: 634 (MH⁺).

[0748] 102h.[2-{2-(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1-(R)-(4-methoxybenzyl)-2-oxo-ethylamino]-aceticacid ethyl ester hydrochloride

[0749] 0.150 g of{[2-{2-(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl{-1-(R)-(4-methoxybenzyl)-2-oxo-ethyl]-tert.-butoxycarbonyl-amino}-aceticacid ethyl ester were treated with 5 mL of HCl in dioxane (5M solution)and the mixture was stirred for 3 h at ambient temperature. The solventwas distilled off and the remainig white solid was washed with diethylether and dried. Yield: 0.100 g (80%); (+)-APCI-MS. 534 (MH⁺).

Example 103 N-(7-amino-thieno[2,3c]pyridin-2-ylmethyl)1-(propyloxycarbonylmethylamino-2(R)-cyclohexylmethyl-acetyl)-prolinamidehydrochloride

[0750] 103a. 7-azido-thieno[2,3c]pyridine

[0751] To a stirred solution of 2 g of 7-chloro-thieno[2,3c]pyridine(U.S. Pat. No. 3,663,559) in 25 mL of dimethylsulfoxide was added 15.6 gof sodium azide and the reaction mixture was heated at 135° C. for 18hours. After this period the reaction mixture was allowed to cool toroom temperature and ethyl acetate and aqueous 1N sodium hydroxide wereadded. The ethyl acetate layer was separated, washed with aqueous 1Nsodium hydroxide (twice) and brine, dried over magnesium sulphate andconcentrated to give 1.65 g of 7-azido-thieno[2,3c]pyridine. MS ESI*:177 (M+H) p 103b. 7-amino-thieno[2,3c]pyridine

[0752] To a stirred solution of 2.1 g of 7-azido-thieno[2,3c]pyridine in250 mL of ethanol was added 1 mL of hydrochloric acid (37%) and 13.4 gof tin(II) chloride dihydrate. The reaction mixture was heated at refluxtemperature for two hours and then concentrated. To the residue wasadded ethyl acetate and a saturated aqueous sodium hydrogen carbonatesolution. After stirring for 30 minutes the mixture was filtered. Theethyl acetate layer of the filtrate was separated, washed with asaturated aqueous sodium hydrogen carbonate solution and brine, driedover magnesium sulphate and concentrated to give 1.65 g of7-amino-thieno[2,3c]pyridine

[0753] TLC: Rf=0.5, dichloromethane/methanol=9/1 v/v on silica.

[0754] 103c. N-[2-(azidomethyl)thieno[2,3c]pyridin-7-yl]benzamide

[0755] The procedures described in examples 1e, 1f, 1g and 73a were usedto convert 7-amino-thieno[2,3c]pyridine intoN-[2-(azidomethyl)thieno[2,3c]pyridin-7-yl]benzamide. Yield: 40%. MSESI*: 310 (M+H)

[0756] 103d. 7-amino-2-(aminomethyl)thieno[2,3c]pyridine

[0757] To a stirred solution of 0.5 g ofN-[2-(azidomethyl)thieno[2,3c]pyridin-7-yl] benzamide in 16 mL of ethylacetate and 24 mL of ethanol was added 3 mL of hydrochloric acid (37%)and 3.6 g of tin(II) chloride dihydrate. The reaction mixture was heatedat reflux temperature for 8 hours and an additional 1 mL of hydrochloricacid (37%) and 1 g of tin(II) chloride dihydrate were added. Afterheating for 17 hours an additional 1 g of tin(II) chloride dihydrate wasadded and the reaction mixture was heated at reflux temperature foranother 6 hours. Then the reaction mixture was concentrated. To theresidue were added ethyl acetate and water and after stirring for 10minutes the mixture was filtered. The filtrate was extracted twice with2N hydrochloric acid. The combined hydrochloric acid extracts were madebasic (pH 10) using aqueous sodium hydroxide and four times extractedwith dichloromethane. The combine dichloromethane extracts were driedover magnesium sulphate and concentrated to give 0.2 g of the titlecompound. MS ESI*: 180 (M+H)

[0758] 103e. N-(7-amino-thieno[2,3c]pyridin-2-ylmethyl)1-(propyloxycarbonylmethylamino-2(R)-cyclohexylmethyl-acetyl)-prolinamidehydrochloride

[0759] A TBTU coupling (procedure described in example 73) of7-amino-2-(aminomethyl)thieno[2,3c]pyridine andN-Boc-N-(1-propyloxycarbonylmethyl)-D-Cha-Pro-OH and subsequently TFAdeprotection gave the TEA salt of the title compound. This TFA salt wasdissolved in dichloromethane, washed with aqueous sodiumhydrogencarbonate, dried over magnesium sulphate and concentrated togive the free base. This free base was dissolved in t-butanol/water,hydrochloric acid was added and lyophilisation afforded the titlecompound. Yield :47%, MS ESI+: 530 (M+H).

Example 104(2-{2-(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-oxo-1-(R)-phenyl-ethyl)-carbamicacid methyl ester (104d)

[0760] 104a. (R)-Methoxy-carbonylamino-phenyl-acetic acid

[0761] Starting with 4.0 g of (D)-phenylglycine gave 2.0 g (35%) of thetitle compound as a white solid using the procedures described inexample 91a. M.p. 122-125° C.; (−)-APCI-MS: 208 ([M−H]⁻).

[0762] 104b.1-[(R)-Methoxycarbonylamino-phenyl-acetyl]-pyrrolidine-2-(S)-carboxylicacid benzyl ester

[0763] Reaction of 1.0 g of (R)-methoxycarbonylamino-phenyl-acetic acidand 1.16 g of (L)-proline benzyl ester hydrochloride according to theprocedure described in example 91b gave 0.59 g (31%) of the titlecompound as a white solid and 0.28 g (15%) of1-[(S)-methoxycarbonylamino-phenyl-acetyl]-pyrrolidine-2-(S)-carboxylicacid benzyl ester (+)-APCI-MS: 397 (MH⁺).

[0764] 104c.1-[(R)-Methoxycarbonylamino-phenyl-acetyl]-pyrrolidine-2-(S)-carboxylicacid

[0765] 1.5 g of1-[(R)-methoxycarbonylamino-phenyl-acetyl]-pyrrolidine-2-(S)-carboxylicacid benzyl ester were dissolved in a mixture of 15 mL of methanol and15 mL of tetrahydrofuran and hydrogenated for 3 h at room temperatureusing 0.4 g palladium on charcoal (10%) as catalyst. Filtration followedby concentration in vacuo affords 0.45 g (100%) of the title compound asa white solid. (+)-APCI-MS: 307 (MH⁺).

[0766] 104d.(2-{2-(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-oxo-1-(R)-phenyl-ethyl)-carbamicacid methyl ester

[0767] Using the procedure described for example 91d 0.36 g of1-[(R)-methoxycarbonylamino-phenyl-acetyl]-pyrrolidine-2-(S)-carboxylicacid was converted into 040 g (74%) of the title compound. (+)-APCI-MS:462 (MH⁺).

Example 105 (2-{2-(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-oxo-1-(S)-phenyl-ethyl)-carbamicacid methyl ester (105b)

[0768] 105a.1-[(S)-Methoxycarbonylamino-phenyl-acetyl]-pyrrolidine-2-(S)-carboxylicacid

[0769] 0.28 g of1-(S)-methoxycarbonylamino-phenyl-acetyl]-pyrrolidine-2-(S)-carboxylicacid benzyl ester were dissolved in a mixture of 15 mL of methanol and15 mL of tetrahydrofuran and hydrogenated for 3 h at room temperatureusing 0.2 g palladium on charcoal (10%) as catalyst. Filtration followedby concentration in vacuo affords 0.21 g (100%) of the title compound asa white solid. (+)-APCI-MS: 307 (MH⁺).

[0770] 105b.(2-{2-(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-oxo-1-(S)-phenyl-ethyl)-carbamicacid methyl ester

[0771] Using the procedure described for example 91d 0.110 g of1-[(S)-methoxycarbonylamino-phenyl-acetyl]-pyrrolidine-2-(S)-carboxylicacid was converted into 0.58 g (36%) of the title compound. (+)-APCI-MS:462 (MH⁺).

Example 106(2-{2-(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1-(R)-cyclohexyl-2-oxo-ethyl)-carbamicacid methyl ester (106d)

[0772] 106a. (R)-Methoxy-carbonylamino-cyclohexyl-acetic Acid

[0773] A mixture of (D)-cyclohexylglycine (6.0 g) in 76.3 mL of 1 N NaOHand 80 mL of dichloromethane was cooled in an ice bath, 3.1 mL of methylchloroformate were added dropwise and the resulting mixture was stirredfor 16 h at room temperature. After adding aqueous NaOH (pH 10) theorganic layer was discarded. KHSO₄ was added (pH 3) and the aqueoussolution was extracted with ethyl acetate. Evaporation of the solvent invacuo gave 2.5 g (30%) of the title compound as a colorless oil.(−)-APCI-MS 214 ([M−H]⁻).

[0774] 106b.1-[(R)-Methoxycarbonylamino-cyclohexyl-acetyl]-pyrrolidine-2-(S-carboxylic acid benzyl ester

[0775] Reaction of 1.24 g of (R)-methoxycarbonylamino-cyclohexyl-aceticacid and 1.40 g of (L)-proline benzyl ester hydrochloride using theprocedure described for example 91b gave 1.17 g (50%) of the titlecompound as a colorless oil. (+)-APCI-MS: 403 (MH⁺).

[0776] 106c.1-[(R)-methoxycarbonylamino-cyclohexyl-acetyl]-pyrrolidine-2-(S)-carboxylicacid

[0777] 1.17 g1-[(R)-methoxycarbonylamino-cyclohexyl-acetyl]-pyrrolidine-2-(S)-carboxylicacid benzyl ester were dissolved in 90 mL of methanol and hydrogenatedfor 3 h at room temperature using 0.8 g palladium on charcoal (10%) ascatalyst. Filtration followed by concentration in vacuo affords 0.80 g(89%) of the title compound as a colorless oil. (−)-APCI-MS: 311([M−H]⁻).

[0778] 106d.(2-{2-(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-oxo-1-(R)-cyclohexyl-ethyl)-carbamicacid methyl ester

[0779] A mixture of1-[(R)-methoxycarbonylamino-cyclohexyl-acetyl]-pyrrolidine-2-(S)-carboxylicacid (0.410 g), 6-aminomethyl-isoquinolin-1-ylamine (0.227 g), N-methylmorpholine (0.58 mL) and 0.423 g of2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate(TBTU) in 10 mL of dry N,N-dimethyl formamide was stirred for 3 h atambient temperature. The mixture was concentrated and the remainingresidue was purified by HPLC (RP-18; H₂O/CH₃OH 95/5→0/100) to give 0.350g (58%) of the title compound as a white solid. (+)-APCI-MS: 468 (MH⁺).

Example 1071-(2-(R)-Methanesulfonylamino-3-phenyl-propionyl)-pyrrolidin-2-(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide (107f)

[0780] 107a. (D)-Phenylalanine methyl ester hydrochloride

[0781] (D)-Phenylalanine (25.0 g) was dissolved in 450 mL of methanoland 22 mL thionyl chloride were slowly added at −20° C. The mixture wasrefluxed for 1 h, followed by evaporation of the volatiles to give 31.2g (97%) of the title compound as a white solid. M.p. 155-163° C.

[0782] 107b. 2-(R)-Methanesulfonylamino-3-phenyl-propionic acid methylester

[0783] To a mixture of (D)-phenylalanine methyl ester hydrochloride(1.10 g) and 2.6 mL of ethyl diisopropylamine in 20 mL ofdichloromethane was added a solution of methanesulfonic anhydride (0.87g) in 20 mL of dichloromethane and the resulting mixture was stirred for1 h. Extraction with 2 N HCl followed by removal of the dichloromethanein vacuo gave 1.70 g of the title compound as a yellow oil

[0784] 107c. 2-(R)-Methanesulfonylamino-3-phenyl-propionic acid

[0785] A mixture of 2-(R)-methanesulfonylamino-3-phenyl-propionic acidmethyl ester (0.90 g) and 0.10 g of LiOH in 10 mL of methanol and 1 mLof water was stirred for 16 h at ambient temperature. KHSO₄ was added,the volatiles were pumped off followed by addition of water, extractionwith dichloromethane and removal of the solvent. Yield: 0.70 g (83%) ofthe title compound as a colorless oil. (−)-APCI-MS: 242 ([M−H]⁻).

[0786] 107d.1-(2-(R)-Methanesulfonylamino-3-phenyl-propionyl)-pyrrolidin-2-(S)-carboxylicacid benzyl ester

[0787] reaction of 0.65 g of2-(R)-methanesulfonylamino-3-phenyl-propionic acid and 0.66 g of(L)-proline benzyl ester hydrochloride using the procedure described forexample xib yielded 0.88 g (75%) of the title compound as a yellow oil.(+)-APCI-MS: 431 (MH⁺).

[0788]107e.1-(2-(R)-Methanesulfonylamino-3-phenyl-propionyl)-pyrrolidin-2-(S)-carboxyicacid

[0789] Hydrogenation of 0.87 g of1-(2-(R)-methanesulfonylamino-3-phenyl-propionyl)-pyrrolidin-2-(S)-carboxylicacid benzyl ester according to the procedure described in example 97eafforded 0.60 g (88%) of the title compound as a light yellow solid.(−)-APCI-MS: 339 ([M−H]⁻).

[0790] 107f.1-(2-(R)-Methanesulfonylamino-3-phenyl-propionyl)-pyrrolidin-2-(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide

[0791] Using the procedure described in example 97f 0.140 g of1-(2-(R)-methanesulfonylamino-3-phenyl-propionyl)-pyrrolidin-2-(S)-carboxylicacid was converted into 0.180 g (90%) of the title compound.(+)-APCI-MS: 496 (MH⁺).

Example 1081-(2-(R)-Acetylamino-3-phenyl-propionyl)-pyrrolidin-2-(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide (108c)

[0792] 108a.1-(2-(R)-Acetylamino-3-phenyl-propionyl)-pyrrolidin-2-(S)-carboxylicacid benzyl ester

[0793] Reaction of 0.42 g of (D)-N-acetyl-phenylalanine and of 0.49 g(L)-proline benzyl ester hydrochloride according to the proceduredescribed for example 91b afforded 0.79 g (100%) of the title compoundas a yellow oil. (+)-APCI-MS: 395 (MH⁺).

[0794] 108b.1-(2-(R)-Acetylamino-3-phenyl-propionyl)-pyrrolidin-2-(S)-carboxylicacid

[0795] Hydrogenation of 0.75 g of1-(2-(R)-acetylamino-3-phenyl-propionyl)-pyrrolidin-2-(S)-carboxylicacid benzyl ester using the procedure described in example 97e afforded0.56 g (96%) of the title compound as a yellow solid. (+)-APCI-MS: 305(MH⁺).

[0796] 108c.1-(2-(R)-Acetylamino-3-phenyl-propionyl)-pyrrolidin-2-(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide

[0797] Using the procedure described for example 97f 0.125 g of1-(2-(R)-acetylamino-3-phenyl)-propionyl)-pyrrolidin-2-(S)-carboxylicacid was converted into 0.90 g (47%) of the title compound. (+)-APCI-MS:460 (MH⁺)

Example 1091-[2-(R)-(3-Ethyl-ureido)-3-(4-methoxy-phenyl)-propionyl]-pyrrolidin-2-(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide (109h)

[0798] 109a.N-tert.-Butoxycarbonyl-2-(R)-amino-3-(4-methoxyphenyl)-propionic acidbenzyl ester)

[0799] (D)-4-Methoxy-phenylalanine benzylester hydrochloride (4.3 g) wasdissolved in 20 mL of water and a solution of di-tert-butyl dicarbonate(3.5 g) in 20 mL of dioxane was added. The mixture was allowed to stirfor 16 h while pH 9.5 was maintained by continious addition of theappropriate amounts of 0.5 N NaOH. The dioxane was distilled offfollowed by extraction with dichloromethane and removal of the solventto give 5.2 g (100%) of the title compound as a colorless oil.

[0800] 109b.N-tert.-Butoxycarbonyl-2-(R)-amino-3-(4-methoxyphenyl)-propionic acid

[0801] Hydrogenation of 5.4 g ofN-tert.-butoxycarbonyl-2-(R)-amino-3-(4-methoxyphenyl)-propionic acidbenzyl ester according to the procedure described in example 97eafforded 3.6 g (90%) of the title compound as a yellow oil. (−)-APCI-MS.294 ([M−H]⁻).

[0802] 109c.1-[(N-tert.-Butoxycarbonyl-2-(R)-amino-3-(4-methoxyphenyl)-propionyl]-pyrrolidin-2-(S)-carboxylicacid benzyl ester

[0803] Reaction of 3.5 g ofN-tert.-Butoxycarbonyl-2-(R)-amino-3-(4-methoxyphenyl)-propionic acidand 2.9 g of (L)-proline benzyl ester hydrochloride according to theprocedure described for example 91b gave 3.0 g (52%) of the titlecompound as a yellow oil. (+)-APCI-MS: 505 (MNa⁺).

[0804] 109d.1-[2-(R)-Amino-3-(4-methoxyphenyl)-propionyl]-pyrrolidin-2-(S)-carboxylicacid benzyl ester hydrochloride

[0805]1-[(N-tert.-butoxycarbonyl)-2-(R)-amino-3-(4-methoxyphenyl)-propionyl]-pyrrolidin-2-(S)-carboxylicacid benzyl ester (2.8 g) were treated with 50 mL of HCl in dioxane (5Msolution) and the mixture was stirred for 4 h at ambient temperature.The solvent was distilled off and the remainig white solid was washedwith diethyl ether and dried. Yield: 2.3 g (96%); (+)-APCI-MS: 383(MH⁺).

[0806] 109e.1-[2-(R)-Isocyanato-3-(4-methoxyphenyl)-propionyl]-pyrrolidin-2-(S)-carboxylicacid benzyl ester

[0807]1-[2-(R)-Amino-3-(4-methoxyphenyl)-propionyl]-pyrrolidin-2-(S)-carboxylicacid benzyl ester hydrochloride (2.8 g) was dissolved in 30 mL ofdichloromethane followed by addition of 30 mL of NaHCO₃ solution (1M)under vigorous stirring. Triphosgene (0.6 g) in 6 mL of dichloromethanewas added at 0° C. and the mixture was kept stirring for 0.25 h. Theorganic layer was separated and the solvent removeed in vacuo to yield1.20 g (98%) of the title compound as a yellow oil. (+)-APCI-MS: 409(MH⁺).

[0808] 109f.1-[2-(R)-(3-Ethyl-ureido)-3-(4-methoxyphenyl)-propionyl]-pyrrolidin-2-(S)-carboxylicacid benzyl ester

[0809]1-[2-(R)-Isocyanato-3-(4-methoxyphenyl)-propionyl]-pyrrolidin-2-(S)-carboxylicacid benzyl ester (0.60 g) was dissolved under nitrogene in 20 mL ofdichloromethane and ethyl amine was allowed to pass through at −20° C.for 5 minutes. The mixture was kept stirring for 16 h. The volatileswere pumped off and the remaining residue was purified by silicachromatography (dichloromethane/methanol 80/20) to give 0.29 g (42%) ofthe title compound as a colorless oil. (+)-APCI-MS: 454(MH⁻).

[0810] 109g. 1-[2-(R)-(3-Ethyl-ureido)-3-(4-methoxyphenyl)-propionyl]-pyrrolidin-2-(S)-carboxylicacid

[0811] Hydrogenation of 0.290 g of1-[2-(R)-(3-ethyl-ureido)-3-(4-methoxyphenyl)-propionyl]-pyrrolidin-2-(S)-carboxylicacid benzyl ester using the procedure described in example 97e afforded0.190 g (82%) of the title compound as a white solid. (−)-APCI-MS: 262([M−H]⁻).

[0812] 109h.1-[2-(R)-(3-Ethyl-ureido)-3-(4-methoxvyphenyl)-propionyl]-pyrrolidin-2-(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide

[0813] Using the procedure described for example 97f 0.190 g of1-[2-(R)-(3-ethyl-ureido)-3-(4-methoxyphenyl)-propionyl]-pyrrolidin-2-(S)-carboxylicacid was converted into 0.180 g (69%) of the title compound.(+)-APCI-MS. 519 (MH⁺).

Example 110 N-(1-amino-isoquinolin-6-ylmethyl)1-(2-acetylamino-4-methyl-6-dimethyl-heptanoyl-prolinamide (110d)

[0814] 110a N-(1-Amino-isoquinolin-6-ylmethyl)-prolinamide

[0815] 5.00 g of 1-Amino-6-aminomethyl-isoquinoline and 6.20 gBoc-L-proline were dissolved in 88 mL DMF. 10.6 g of TBTU was addedfollowed by a dropwise addition of N-methylmorpholine. Stirring wascontinued for 1 h at ambient temperature and the solvent was removedi.vac. The residue was extracted with 5% aqueous NaHCO3 solution andwith ethyl acetate. The organic layer was separated and dried (NaSO4).The solvent was removed to give 10.7 g (quant.) ofN-(1-Amino-isoquinolin-6-ylmethyl)-1-tert.butoxycarbonyl-prolinamide asan oil. Without purification, the compound was dissolved in 50 mLdichloromethane and 25.0 mL trifluoroacetic acid was added. Stirring wascontinued for 16 h and the solvent was removed i.vac. The residue wasdissolved in methanol, the solvent was removed i. vac. And the residuewas triturated with diethyl ether. The crystalline material wascollected to give 12.9 g (89%) ofN-(1-amino-isoquinolin-6-ylmethyl)-prolinamide. M.p. 140-142° C.

[0816] 110b.N-(1-Amino-isoquinolin-6-ylmethyl)-1-tert.butoxycarbonyl-azetidin-2(S)-carboxamide

[0817] 2.00 g 1-Tert.butoxycarbonyl-azetidin-2(S)-carboxylic acid, 1.70g 1-amino-6-aminomethyl-isoquinolin, 3.50 g TBTU were dissolved in 40 mLDMF. 3.55 mL N-methylmorpholin was added slowly with stirring. Thesolution was applied to a reversed-phase chromatography column (lenght100 mm, diameter 14 mm; Merck Lichroprep RP-18, 15-25 μ). Elution wasstarted with 25 mL of methanol/water containing 0.3% acetic acid(10:90), then a linear gradient to pure methanol over a period of 75 mLwas applied followed by 50 mL of pure methanol. Fraktions of 5 mL eachwere sampled. The solvent was evaporated to give 3.4 g of the tilecompound. MS: 357

[0818] 110c.N-(1-Amino-isoquinolin-6-ylmethyl)-azetidin-2(S)-carboxamide

[0819] 10 mL Trifluoroacetic acid was added to a solution of 2,4 g ofN-(1-amino-isoquinolin-6-ylmethyl)-1-tert.butoxycarbonyl-azetidin-2(S)-carboxamidein 5.0 mL dichloromethane. The solvent was removed i.vac to give 2.1 gof the title comound as trifluoroacetate salt. MS: 371.

[0820] 110c. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-acetylamino-4-methyl-6-dimethyl-heptanoyl)-prolinamide

[0821] 125 mg of N-(1-amino-isoquinolin-6-ylmethyl)-prolinamide wasdissolved in 2 mL DMF, 60 mg of2-acetylamino-4-methyl-6-dimethyl-heptanoic acid, 80 mg of TBTU and 75mg of N-methylmorpholine were added. The solution was stirred for 1 h atambient temperature. The solution was applied to a reversed-phasechromatography column (lenght 100 mm, diameter 14 mm; Merck LichroprepRP-18, 15-25 μ). Elution was started with 25 mL of methanol/watercontaining 0.3% acetic acid (10:90), then a linear gradient to puremethanol over a period of 75 mL was applied followed by 50 mL of puremethanol. Fraktions of 5 mL each were sampled. The compound was detectedin fractions 10-12 by mass spektra. The solvent was removed i.vac. togive 105 mg (87%) of the title compound as amorphous solid. MS: 482.

Example 111

[0822] The following compounds were prepared analogously to Example 110:

[0823] 111a. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-acetylamino-2-(3,4-ethylenedioxy-phenyl)-acetyl)-prolinamide

[0824] Yield 38%. MS: 504.

[0825] 111b. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-acetylamino-2-(3-methoxy-phenyl)-acetyl)-prolinamide

[0826] Yield 27%. MS: 476

[0827] 111c. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-acetylamino-2-(2-methoxy-phenyl)-acetyl)-prolinamide

[0828] Yield 78%. MS: 476

[0829] 111d. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-acetylamino-2-(4-fluoro-phenyl)-acetyl)-prolinamide

[0830] Yield 54%. MS: 464

[0831] 111e. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-acetylamino-2-(thiophen-3-yl)-acetyl)-prolinamide

[0832] Yield 86%. MS: 452

[0833] 111f. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-acetylamino-2-(naphthalene-2-yl)-acetyl)-prolinamide

[0834] Yield 51%. MS: 496.

[0835] 111g. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-acetylamino-2-(4-methoxycarbonyl-phenyl)-acetyl)-prolinamide

[0836] Yield 39%. MS: 504.

[0837] 111h. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-acetylamino-2-(4-cyano-phenyl)-acetyl)-prolinamide

[0838] Yield 21%. MS: 471.

[0839] 111i. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-acetylamino-2-(4-trifluoromethyl-phenyl)-acetyl)-prolinamide

[0840] Yield 29%. MS: 514.

[0841] 111j. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-acetylamino-2-(2-chloro-phenyl)-acetyl)-prolinamide

[0842] Yield 78%. MS: 481.

[0843] 111k. N-(1-amino-isoquinolin-6-ylmethyl) 1-(2-acetylamino-2-(3-chloro-phenyl)-acetyl)-prolinamide

[0844] Yield 34%. MS: 481.

[0845] 111l. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-acetylamino-2-(4-chloro-phenyl)-acetyl)-prolinamide

[0846] Yield 40%. MS: 481.

[0847] 111m. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-acetyl-methyl-amino-2-cyclohexyl-acetyl)-prolinamide

[0848] Yield 71%. MS: 466.

[0849] 111n. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-acetylamino-2-cyclohexyl-acetyl)-prolinamide

[0850] Yield 64%. MS: 452.

[0851] 111o. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-acetoxy-2-(4-trifluormethyl-phenyl)-acetyl)-prolinamide

[0852] Yield 57%. MS: 515.

[0853] 111p. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-(2-methoxyphenyl)-sulfonylamino-2(R)-phenyl-acetyl)-prolinamide

[0854] Yield 68%. MS: 574.

[0855] 111q. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-benzylcarbonylamino-2(R)-phenyl-acetyl)-prolinamide

[0856] Yield 54%. MS: 522.

[0857] 111r. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-formylamino-2(R)-phenyl-acetyl)-prolinamide

[0858] Yield 95%. MS: 432.

[0859] 111s. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-benzylsulfonylamino-2(R)-phenyl-acetyl)-prolinamide

[0860] Yield 57%. MS: 558.

[0861] 111t. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-methylsulfonylamino-2(R)-phenyl-acetyl)-prolinamide

[0862] Yield 67%. MS: 482.

[0863] 111u. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-ethylsulfonylamino-2(R)-phenyl-acetyl)-prolinamide

[0864] Yield 59%. MS: 496.

[0865] 111v. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-acetylamino-2-(4-bromo-phenyl)-acetyl)-prolinamide

[0866] Yield 63%. MS: 526.

[0867] 111w. N-(1-amino-isoquinolin-6-ylmethyl)1-(9-hydroxy-fluorenyl-9-yl)-carbonyl-prolinamide

[0868] Yield 24%. MS: 479.

[0869] 111x. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-acetoxy-2-(4-chlorophenyl)-acetyl)-prolinamide

[0870] Yield 62%. MS: 481.

[0871] 111y. N-1-amino-isoquinolin-6-ylmethyl)1-(2-acetoxy-2-(4-methoxyphenyl)-acetyl)-prolinamide

[0872] Yield 63%. MS: 477.

[0873] 111z. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-phenylsulfonylamino-2(R)-phenyl-acetyl)-prolinamide

[0874] Yield 89%. MS: 544.

[0875] 111aa. N-(1-amino-isoquinolin-6-ylmethyl)1-(acetoxy-2(R)-cyclohexyl-acetyl)-prolinamide

[0876] Yield 79%. MS: 453.

[0877] 111ab. N-(1-amino-isoquinolin-6-ylmethyl)1-(methoxycarbonylmethylamino-2(R)-cyclohexylmethyl-acetyl)-prolinamide

[0878] Yield 16%. MS: 496.

[0879] 111ac. N-(1-amino-isoquinolin-6-ylmethyl)1-(propyloxycarbonylmethylamino-2(R)-cyclohexylmethyl-acetyl)-prolinamide

[0880] Yield 30%. MS: 524.

[0881] 111ad. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-ethoxycarbonylmethylamino-2(R)-cyclohexylmethyl-acetyl)-prolinamide

[0882] Yield 60%. MS 510.

[0883] 111ae.[(2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-cyclohexylmethyl-2-oxo-ethyl)-tert-butoxycarbonylmethyl-amino]-aceticacid tert-butyl ester

[0884] Yield 25%. MS: 652.

[0885] 111af.[(2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}(R)-cyclohexylmethyl-2-oxo-ethyl)-methyl-amino]-aceticacid tert-butyl ester

[0886] Yield 20%. MS: 552.

[0887] 111ag.1-((R)-Phenyl-propionylamino-acetyl)-pyrrolidine-2(S)-carboxylic acid(1-amino-isoquinolin-6-ylmethyl)-amide

[0888] Yield 63%. MS: 460.

[0889] 111ah.1-((R)-Acetylamino-phenyl-acetyl)-pyrrolidine-2(S)-carboxylic acid(1-amino-isoquinolin-6-ylmethyl)-amide

[0890] Yield 61%. MS: 446.

[0891] 111ai.1-((R)-Benzoylamino-phenyl-acetyl)-pyrrolidine-2(S)-carboxylic acid(1-amino-isoquinolin-6-ylmethyl)-amide

[0892] Yield 40%. MS: 508.

[0893] 111aj.(2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-oxo-1(R)-phenyl-ethyl)-carbamicacid tert-butyl ester

[0894] Yield 68%. MS: 504.

[0895] 11ak. 1-(3,3-Diphenyl-propionyl)-pyrrolidine-2(S)-carboxylic acid(1-amino-isoquinolin-6-ylmethyl)-amide

[0896] Yield 69%. MS: 479.

[0897] 111al.1-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl-pyrrolidine-1-carbonyl}-octahydro-isoquinoline-2-carboxylicacid methyl ester

[0898] Yield 74%. MS: 494.

[0899] 111am. Acetic acid2-{2(S)-[(1-amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin1-yl}-2-oxo-1(R)-phenyl-ethylester

[0900] Yield 69%. MS: 447.

[0901] 111an.[2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-(4-fluoro-benzyl)-2-oxo-ethyl]-carbamicacid tert-butyl ester

[0902] Yield 83%. MS: 536.

[0903] 111ao.[2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-(4-methoxy-benzyl)-2-oxo-ethyl-carbamicacid tert-butyl ester

[0904] Yield 76%. MS: 548.

[0905] 111ap.(2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-benzyl-2-oxo-ethyl)-carbamicacid tert-butyl ester

[0906] Yield 81%. MS: 518.

[0907] 111aq.(2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-benzyl-2-oxo-ethyl)-carbamicacid benzyl ester

[0908] Yield 82%. MS: 552.

[0909] 111ar. Acetic acid2-{2(S)-[(1-amino-isoquinolin-6-ylmethyl)-carbamoyl]-azetidin-1-yl}-2-oxo-1(R)-phenyl-ethylester

[0910] Yield 23%. MS: 433.

[0911] 111as.[2-{2(S)-[(1-amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-(4-methoxy-benzyl)-2-oxo-ethylamino]-aceticacid.

[0912] Yield 91%. MS: 506.

[0913] 111at.1-(3-Cyclohexyl-2(R)-ethanesulfonylamino-propionyl)-pyrrolidine-2(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide

[0914] Yield 50%. MS: 516.

[0915] 111av.(2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-azetidin-1-yl}-2-oxo-1(R)-phenyl-ethyl)-carbamicacid ethyl ester

[0916] Yield 43%. MS: 462

Example 112 N-(1-amino-isoquinolin-6-ylmethyl)1-(2-hydroxycarbonylmethylamino-2(R)-cyclohexylmethyl-acetyl)-prolinamide

[0917] To a solution of 135 mg ofN-(1-amino-isoquinolin-6-ylmethyl)-1-(propyloxycarbonylmethylamino-2(R)-cyclohexylmethyl-acetyl)-prolinamide in 4.0 mLdimethoxyethane was added 2.0 mL of 2 M aqueous lithium hydroxidesolution was added and stirring was continued for 3 h at ambienttemperature. The solvent was removed i.vac., methanol was added and thecompound was purified by reversed-phase column chromatography asdescribed in example 110c. Yield: 100 mg (81%). MS: 482

Example 113

[0918] The following compounds were prepared using the proceduredescribed in example 112:

[0919] 113a. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-hydroxy-2(R)-cyclohexyl-acetyl)-prolinamide

[0920] Yield (84%) MS: 411.

[0921] 113b. 1-((R)-Hydroxy-phenyl-acetyl)-pyrrolidine-2(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide

[0922] Yield 93%. MS: 404.

Example 114 1-((R)-Amino-phenyl-acetyl)-pyrrolidine-2(S)-carboxylic acid(1-amino-isoquinolin-6-ylmethyl)-amide

[0923] Trifluoroacetic acid (3.0 mL) was added to 150 mg of(2-{2(S)-[(1-amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-oxo-1(R)-phenyl-ethyl)-carbamicacid tert-butyl ester (prepared according to the procedures described inexample 110) in 3.0 mL dichloromethane. Stirring was continued for 2 hat ambient temperature. The solvent was removed i.vac, the residue wasdissolved in methanol and applied to a reversed-phase chromatographycolumn (lenght 100 mm, diameter 14 mm; Merck Lichroprep RP-18, 15-25 μ).Elution was started with 25 mL of methanol/water containing 0.3% aceticacid (10:90), then a linear gradient to pure methanol over a period of75 mL was applied followed by 50 mL of pure methanol. Fractions of 5 mLeach were sampled. The solvent was removed to give 75 mg (62%) of thetitle compound. MS: 404.

Example 115

[0924] The following compounds were prepared using the proceduredescribed in example 114:

[0925] 115a.1-[2(R)-Amino-3-(4-fluoro-phenyl)-propionyl]-pyrrolidine-2(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide

[0926] Yield 99%. MS: 436.

[0927] 115b.1-[2(R)-Amino-3-(4-methoxy-phenyl)-propionyl]-pyrrolidine-2(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide

[0928] Yield 90%. MS: 446.

[0929] 115c.1-(2(R)-Amino-3-phenyl-propionyl)-pyrrolidine-2(S)-carboxylic acid(1-amino-isoquinolin-6-ylmethyl-amide

[0930] Yield 94%. MS: 418.

[0931] 115d.[2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl{-1(R)-(4-chloro-benzyl)-2-oxo-ethylamino]-aceticacid

[0932] Yield 84%. MS: 510

[0933] 115e.(2-{2-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-oxo-1-phenyl-ethylamino)-aceticacid

[0934] Yield 95%. MS: 462.

[0935] 115f.(2-{2-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1-cyclohexyl-2-oxo-ethylamino)-aceticacid

[0936] Yield 80%. MS: 468.

[0937] 115g.(2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-azetidin-1-yl}-1(R)-cyclohexyl-2-oxo-ethylamino)-aceticacid

[0938] Yield 49%. MS: 454.

[0939] 115h.1-[2-Amino-3(R)-(4-chloro-phenyl)-propionyl]-pyrrolidine-2(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide

[0940] Yield 36%. MS: 452.

[0941] 115i.[2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-(4-fluoro-benzyl)-2-oxo-ethylamino]-aceticacid

[0942] Yield 25%. MS: 494.

Example 116(2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-benzyl-2-oxo-ethyl)-carbamicacid benzyl ester

[0943] 12.5g (10 mmol) of the aminomethyl-polystyrene resine (Bachem) in100 mL dry THF was shaken for 1 h, 4.6 g of 4-aminosulfonyl-butyricacid, 4.5 mL of diisopropylcarbodiimid and 3.8 g of hydroxybenzotriazolewere added and shaken at ambient temperature for 16 h. The resin wasfiltered, washed with THF, DMF, methanol and ether, and dried at 50° C.to give 13.7 g of dry resin.

[0944] This resin was shaken in 100 mL THF for 1 h at ambienttemperature. To a solution of 3.2 g of N-Boc-proline in 30 mL dry THFwas added 2.5 g of carbonyldiimidazole and strirred for 30 min, thenrefluxed for 30 min, and cooled to ambient temperature. This solutionwas added to the slurry of the resin in THF, followed after 10 min by2.7 mL of DBU. Shaking was continued for 16 h at ambient temperature,the resin was filtered, washed with acetic acid, DMF, methanol andether, and dried i.vac. at 50° C. for 3 h to give 13.3 g of dry resin.

[0945] The resin was shaken in 150 mL dichloromethane for 1 h at ambienttemperature. It was filtered, 50 mL of trifluoroacetic acid and 2.5 mLwater was added, shaken for 45 min at ambient temperature, filtered,washed with dry dichloromethane and methanol, and dried i.vac. at 50° C.for 16 h.

[0946] 150 mg of this dried resin was shaken with 2.4 mL of a 1 Nsolution of N-methyl-morpholine in DMF for 20 min, filtered, shaken with2.4 mL DMF, and filtered. 0.4 mL of a 1 M solution ofZ-(R)-phenylalanine in DMF was added, followed by 1.0 mL of a solutionof a 0.4 M solution of TBTU in DMF and by 1.0 mL of a 1 M solution ofNMM in DMF. The resin was shaken for 3.5 h at 40° C., filtered andwashed three times with 2.4 mL each of DMF. Again, 0.4 mL of a 1 Msolution of Z-(R)-phenylalanine in DMF was added, followed by 1.0 mL ofa solution of a 0.3 M solution of TBTU in DMF and by 1.0 mL of a 1 Msolution of NMM in DMF. The resin was shaken for 3.5 h at 40° C.,filtered and washed three times with 2.4 mL each of DMF, three timeswith 2.4 mL each of dichloromethane, once with 3.0 mL methanol, andtwice with 2.4 mL each of DMF.

[0947] 1.4 mL Of a 0.25 M solution of DIPEA in DMF followed by 1.6 mL ofa 1M solution of iodoacetonitrile in DMF was added, and the resin wasshaken at 40° C. for 4.5 h. The resin was filtered, washed three timeswith 2.4 mL each of DMF. Then, 1.4 mL of a 0.25 M solution of DIPEA inDMF followed by 1.6 mL of a 1M solution of iodoacetonitrile in DMF wasadded, and the resin was shaken at 40° C. for 4.5 h, filtered, washedthree times with 2.4 mL each of DMF, three times with 2.4 mL each ofdichloromethane, once with 3.0 mL methanol, and five times with 2.4 mLeach of dichloromethane.

[0948] 2.0 mL of a 0.05 M solution of6-(aminomethyl)-isoquinoline-1-ylamine in dichloromethane followed by1.0 mL dichloromethane was added to the resin and shaken for 4 h atambient temperature. The solvent was collected by filtration and theresin was washed with 2.0 mL dichloromethane and 2.0 mL methanol. Theorganic phases were combined. The presence of the desired compound wasdetected by mass spectrum, and its purity was determined by TLC. Thesolvent was removed i.vac. to give(2-{2(S)-[(1-amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-benzyl-2-oxo-ethyl)-carbamicacid benzyl ester MS: 552

Example 117

[0949] The following compounds were prepared using the proceduredescribed in example 116:

[0950] 117a.1-{1(S)-[(4-Amino-thieno[3.2-c]pyridin-2-ylmethyl)-carbamoyl]-2-naphthalen-1-yl-ethylcarbamoyl{-octahydro-isoquinoline-2-carboxylicacid methyl ester

[0951] MS: 600

[0952] 117b.1-{2(S)-[(4-Amino-thieno[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-pyrrolidine-1-carbonyl{-octahydro-isoquinoline-2-carboxylicacid methyl ester

[0953] MS 500

[0954] 117c.(2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-cyclohexylmethyl-2-oxo-ethyl)-carbamicacid benzyl ester

[0955] MS: 558

[0956] 117d.(2-{2(S)-[(4-Amino-thieno[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-benzyl-2-oxo-ethyl)-carbamicacid benzyl ester

[0957] MS: 558

[0958] 117e.(2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-azetidin-1-yl{-1(R)-benzyl-2-oxo-ethyl)-carbamicacid benzyl ester

[0959] MS: 538

[0960] 117f.(2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-piperidin-1-yl}-1(R)-benzyl-2-oxo-ethyl)-carbamicacid benzyl ester

[0961] MS: 560

[0962] 117g.(2-{2(S)-[(1-Amino-isoquinolin-7-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-benzyl-2-oxo-ethyl)-carbamicacid benzyl ester

[0963] MS: 552

[0964] 117h.(2-{2(S)-[(1-Amino-isoquinolin-7-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-cyclohexylmethyl-2-oxo-ethyl)-carbamicacid benzyl ester

[0965] MS: 558

[0966] 117i.(2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-azetidin-1-yl}-1(R)-cyclohexylmethyl-2-oxo-ethyl)-carbamicacid benzyl ester

[0967] MS: 544

[0968] 117j.(2-{2(S)-[(1-Amino-isoquinolin-7-ylmethyl)-carbamoyl]-azetidin-1-yl}-1(R)-cyclohexylmethyl-2-oxo-ethyl)-carbamicacid benzyl ester

[0969] MS: 544

[0970] 117k.[2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-piperidin-1-yl}-1(R)-(4-methoxy-benzyl)-2-oxo-ethyl]-carbamicacid tert-butyl ester

[0971] MS: 562

[0972] 117l.1-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-azetidine-1-carbonyl}-octahydro-isoquinoline-2-carboxylicacid methyl ester

[0973] MS: 480

[0974] 117m.1-{2(S)-[(1-Amino-isoquinolin-7-ylmethyl)-carbamoyl]-pyrrolidine-1-carbonyl}-octahydro-isoquinoline-2-carboxylicacid methyl ester

[0975] MS: 494

[0976] 117n.1-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-piperidine-1-carbonyl}-octahydro-isoquinoline-2-carboxylicacid methyl ester

[0977] MS: 508

[0978] 117o.(2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-piperidin-1-yl}-2-oxo-1(R-phenyl-ethyl)-carbamicacid tert-butyl ester

[0979] MS: 518

[0980] 117p.1-{1(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-2-naphthalen-1-yl-ethylcarbamoyl}-octahydro-isoquinoline-2-carboxylicacid methyl ester

[0981] MS: 594

[0982] 117q. 1(R)-(Benzoylamino-phenyl-acetyl)-azetidine-2(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide

[0983] MS: 494

[0984] 117r.1(R)-(Benzoylamino-phenyl-acetyl)-pyrrolidine-2(S)-carboxylic acid(1-amino-isoquinolin-7-ylmethyl)-amide

[0985] MS: 508

[0986] 117s.N-(4-Amino-thieno[3.2-c]pyridin-2-ylmethyl)-2(S)-(3,3-diphenyl-propionylamino)-3-naphthalen-1-yl-prolinamide

[0987] MS: 585

[0988] 117t.1(R)-(Acetylamino-phenyl-acetyl)-Dyrrolidine-2(S)-carboxylic acid(1-amino-isoquinolin-7-ylmethyl)-amide

[0989] MS: 446

[0990] 117v.N-(1-Amino-isoquinolin-6-ylmethyl)-3-naphthalen-1-yl-2(S)-(2-phenoxy-acetylamino)-prolinamide

[0991] MS: 505

[0992] 117w.1(R)-(Phenyl-propionylamino-acetyl)-pyrrolidine-2(S)-carboxylic acid(1-amino-isoquinolin-7-ylmethyl)-amide

[0993] MS: 460

[0994] 117x.1-[(R)-(2,2-Dimethyl-propionylamino)-phenyl-acetyl]-pyrrolidine-2(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide

[0995] MS: 488

[0996] 117y.1-[(R)-(2,2-Dimethyl-propionylamino)-phenyl-acetyl]-pyrrolidine-2(S)-carboxylicacid (1-amino-isoquinolin-7-ylmethyl)-amide

[0997] MS: 488

[0998] 117z.(1-{1(S)-[(4-Amino-thieno[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-2-naphthalen-1-yl-ethylcarbamoyl}-2(R)-naphthalen-1-yl-ethyl)-carbamicacid tert-butyl ester

[0999] MS: 674

[1000] 117aa.(1-{1(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-2-naphthalen-1-yl-ethylcarbamoyl}-2(R)-naphthalen-1-yl-ethyl)-carbamicacid tert-butyl ester

[1001] MS: 668

[1002] 117ab.(3(R)-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidine-1-carbonyl}-3,4-dihydro-1H-isoquinolin-2-yl)-aceticacid tert-butyl ester

[1003] MS: 544

[1004] 117ac. Acetic acid2-{2(S)-[(4-amino-furo[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-azetidin-1-yl}-2-oxo-1(S)-phenyl-ethylester

[1005] MS: 423

[1006] 117ad. Acetic acid2-{2(S)-[(4-amino-furo[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-oxo-1(S)-phenyl-ethylester

[1007] MS: 437

[1008] 117ae. Acetic acid2-{2(S)-[(4-amino-furo[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-piperidin-1-yl}-2-oxo-1(S)-phenyl-ethylester

[1009] MS: 451

[1010] 117af. 1-Phenoxyacetyl-pyrrolidine-2(S)-carboxylic acid(1-amino-isoquinolin-6-ylmethyl)-amide

[1011] MS: 405

[1012] 117ag. Acetic acid2-{2(S)-[(1-amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-oxo-1(S)-phenyl-ethylester

[1013] MS: 447

[1014] 117ah. Acetic acid2-{2(S)-[(1-amino-isoquinolin-6-ylmethyl)-carbamoyl]-piperidin-1-yl}-2-oxo-1(S)-phenyl-ethylester

[1015] MS: 461

[1016] 117ai. 1-(3,3-Diphenyl-propionyl)-azetidine-2(S)-carboxylic acid(1-amino-isoquinolin-6-ylmethyl)-amide

[1017] MS: 465

[1018] 117aj. 1(R)-(Methoxy-phenyl-acetyl)-pyrrolidine-2(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide

[1019] MS: 419

[1020] 117ak. 1((S)-Methoxy-phenyl-acetyl)-pyrrolidine-2(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide

[1021] MS: 419

[1022] 117al. 1-((R)-Methoxy-phenyl-acetyl)-tyrrolidine-2(S)-carboxylicacid (4-amino-thieno[3,2-c]pyridin-2-ylmethyl)-amide

[1023] MS: 425

[1024] 117am. 1-((S)-Methoxy-phenyl-acetyl)-pyrrolidine-2(S)-carboxylicacid (4-amino-thieno[3,2-c]pyridin-2-ylmethyl)-amide

[1025] MS: 425

[1026] 117an. Acetic acid9-{2(S)-[(1-amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidine-1-carbonyl}-9H-fluoren-9-ylester

[1027] MS: 521

[1028] 117ao. Acetic acid9-{2(S)-[(1-amino-isoquinolin-6-ylmethyl)-carbamoyl]-azetidine-1-carbonyl}-9H-fluoren-9-ylester

[1029] MS: 507

[1030] 117ap. 1-(3,3-Diphenyl-propionyl)-pyrrolidine-2(S)-carboxylicacid (1-amino-isoquinolin-7-ylmethyl)-amide

[1031] MS: 479

[1032] 117aq. 1-(3,3-Diphenyl-propionyl)-pyrrolidine-2(S)-carboxylicacid (4-amino-thieno[3,2-c]pyridin-2-ylmethyl)-amide

[1033] MS: 485

[1034] 117ar. 1-[(9H-Fluoren-9-yl)-acetyl]-pyrrolidine-2(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide

[1035] MS: 477

[1036] 117as.[2-}2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-(4-chloro-benzyl)-2-oxo-ethyl]-carbamicacid tert-butyl ester

[1037] MS: 553

Example 118

[1038] The procedure described in example 116 was repeated. After theproducts had been isolated, the Boc-protecting groups were removed bytreating the residues with 1 mL of 50% trifluoroacetic acid indichloromethane and stirring for 20 min at ambient temperature. Thesolvent was removed in vacuo and the presence of the desired compoundwas detected by mass spectrum, and its purity was determined by TLC.

[1039] 118a.1-(2(R)-Amino-3-naphthalen-1-yl-propionyl)-pyrrolidine-2(S)-carboxylicacid (4-amino-furo[3,2-c]pyridin-2-ylmethyl)-amide

[1040] MS: 458

[1041] 118b.1-(2(R)-Amino-3-naphthalen-1-yl-propionyl)-azetidine-2(S)-carboxylicacid (4-amino-furor[3,2-c]pyridin-2-ylmethyl)-amide

[1042] MS: 444

[1043] 118c.1-(2(R)-Amino-3-naphthalen-1-yl-propionyl)-azetidine-2(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide.

[1044] MS: 454

[1045] 118d.(2-{2(S)-[(4-Amino-furo[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-cyclohexyl-2-oxo-ethylamino)-aceticacid

[1046] MS: 458

[1047] 118e.(2-{2(S)-[(4-Amino-furo[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-azetidin-1-yl}-1(R)-cyclohexyl-2-oxo-ethylamino)-aceticacid

[1048] MS: 444

[1049] 118f. 1-(2(S)-Benzylamino-propionyl)-pyrrolidine-2(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide

[1050] 118g.(2-{2(S)-[(4-Amino-furo[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-oxo-1(R)-phenyl-ethylamino)-aceticacid

[1051] MS: 452

[1052] 118h.(2-{2(S)-[(4-Amino-furo[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-azetidin-1-yl}-2-oxo-1(R)-phenyl-ethylamino)-aceticacid

[1053] MS: 438

[1054] 118i. 1-(2(R)-Benzylamino-propionyl)-pyrrolidine-2(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide. MS: 432

[1055] 118j.(2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-azetidin-1-yl}-2-oxo-1(R)-phenyl-ethylamino)-aceticacid

[1056] MS: 448

[1057] 118k.(2-{2(S)-[(4-Amino-furo[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-benzyl-2-oxo-ethylamino)-aceticacid

[1058] MS: 466

[1059] 118l.(2-{2(S)-[(4-Amino-furo[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-azetidin-1-yl}-1(R)-benzyl-2-oxo-ethylamino)-aceticacid

[1060] MS: 452

[1061] 118m.(2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-benzyl-2-oxo-ethylamino)-aceticacid

[1062] MS: 476

[1063] 118n.(2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-azetidin-1-yl}-1(R)-benzyl-2-oxo-ethylamino)-aceticacid

[1064] 118o.(2-{2(S)-[(1-Amino-isoquinolin-7-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-benzyl-2-oxo-ethylamino)-aceticacid OBM 14.1688

[1065] MS: 476

[1066] 118p.[2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-azetidin-1-yl}-1(R)-(4-fluoro-benzyl)-2-oxo-ethylamino]-aceticacid

[1067] MS: 480

[1068] 118q.(2-{2(S)-[(4-Amino-thieno[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-azetidin-1-yl}-1(R)-cyclohexyl-2-oxo-ethylamino)-aceticacid

[1069] MS: 460

[1070] 118r.[2-{2(S)-[(4-Amino-thieno[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-azetidin-1-yl}-1(R)-(4-fluoro-benzyl)-2-oxo-ethylamino-aceticacid

[1071] MS: 486

[1072] 118s.[2-{2(S)-[(4-Amino-thieno[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-(4-fluoro-benzyl)-2-oxo-ethylamino-aceticacid

[1073] MS: 500

[1074] 118t(2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-naphthalen-1-ylmethyl-2-oxo-ethylamino)-aceticacid

[1075] MS: 526

[1076] 118u.(2-{2(S)-[(4-Amino-thieno[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-azetidin-1-yl}-1(R)-cyclohexyl-2-oxo-ethylamino)-aceticacid tert-butyl ester

[1077] 118y.(2-{2(S)-[(4-Amino-thieno[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-oxo-1(R)-phenyl-ethylamino)-aceticacid tert-butyl ester

[1078] MS: 524

[1079] 118w.(2-{2(S)-[(4-Amino-thieno[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-azetidin-1-yl}-2-oxo-1(R)-phenyl-ethylamino)-aceticacid tert-butyl ester

[1080] MS: 510

[1081] 118x.(2-{2(S)-[(4-Amino-thieno[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-cyclohexyl-2-oxo-ethylamino)-aceticacid tert-butyl ester

[1082] MS: 530

[1083] 118y.(2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1yl}-1(R)-naphthalen-2-ylmethyl-2-oxo-ethylamino]-aceticacid

[1084] MS: 526

[1085] 118z.(2-{2(S)-[(1-Amino-isoquinolin-7-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-cyclohexyl-2-oxo-ethylamino)-aceticacid

[1086] MS: 468

[1087] 118aa.[2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-azetidin-1-yl}-1-(R)-(4-chloro-benzyl)-2-oxo-ethylamino]-aceticacid

[1088] MS: 497

[1089] 118ab.[2-{2(S)-[(1-Amino-isoquinolin-7-ylmethyl)-carbamoyl]-pyrrolidin-1yl}-1(R)-(4-chloro-benzyl)-2-oxo-ethylamnino]-aceticacid

[1090] MS: 5.1

[1091] 118ac.[2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-azetidin-1-yl}-1(R)-(4-methoxy-benzyl)-2-oxo-ethylamino]-aceticacid

[1092] MS: 492

[1093] 118ad.[2-{2(S)-[(1-Amino-isoquinolin-7-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}1(R)-(4-methoxy-benzyl)-2-oxo-ethylamino]-aceticacid

[1094] MS: 506

[1095] 118ae. 1-Phenylaminoacetyl-azetidine-2(S)-carboxylic acid(4-amino-furo[3,2-c]pyrdin-2-ylmethyl)-amide

[1096] MS: 380

[1097] 118af. 1-Phenylaminoacetyl-azetidine-2(S)-carboxylic acid(1-amino-isoquinolin-6-ylmethyl)-amide

[1098] 118ag. 1-Phenylaminoacetyl-pyrrolidine-2(S)-carboxylic acid(1-amino-isoquinolin-6-ylmethyl)-amide

[1099] MS: 404

[1100] 118ah. 1-Phenylaminoacetyl-piperidine-2(S)-carboxylic acid(1-amino-isoquinolin-6-ylmethyl)-amide

[1101] MS: 418

[1102] 118ai. 1-Phenethylaminoacetyl-azetidine-2(S)-carboxylic acid(4-amino-furo[3,2-c]pyridin-2-ylmethyl)-amide

[1103] MS: 408

[1104] 118aj. 1-Phenethylaminoacetyl-azetidine-2(S)-carboxylic acid(1-amino-isoquinolin-6-ylmethyl)-amide

[1105] MS: 418

[1106] 118ak. 1-Phenethylaminoacetyl-pyrrolidine-2(S)-carboxylic acid(1-amino-isoquinolin-6-ylmethyl)-amide

[1107] MS: 432

[1108] 118al. 1-Phenethylaminoacetyl-piperidine-2(S)-carboxylic acid(1-amino-isoquinolin-6-ylmethyl)-amide

[1109] MS: 446

[1110] 118am. 1-Benzylaminoacetyl-azetidine-2(S)-carboxylic acid (1-amino-isoquinolin-6-ylmethyl)-amide

[1111] MS: 404

[1112] 118an. 1-Benzylaminoacetyl-pyrrolidine-2(S)-carboxylic acid(1-amino-isoquinolin-6-ylmethyl)-amide

[1113] MS: 418

[1114] 118ao. 1-Benzylaminoacetyl-piperidine-2(S)-carboxylic acid(1-amino-isoquinolin-6-ylmethyl-amide

[1115] MS: 432

[1116] 118ap. 1-Cyclopentylaminoacetyl-pyrrolidine-2(S)-carboxylic acid(1-amino-isoquinolin-6-ylmethyl)-amide

[1117] MS: 396

[1118] 118aq.N-(1-Amino-isoquinolin-6ylmethyl)-2(S)-(2-cyclohexylamino-acetylamino)-3-naphthalen-1-yl-prolinamide

[1119] MS: 510

[1120] 118ar.N-(4-Amino-thieno[3,2-c]pyridin-2-ylmethyl)-2(S)-(2-cyclohexylamino-acetylamino)-3-naphthalen-1-yl-prolinamide

[1121] MS; 516

[1122] 118as.2(S)-Amino-N-{1-[(1-amino-isoquinolin-6ylmethyl)-carbamoyl]-2-naphthalen-1-yl-ethyl}-4-phenyl-butyramide

[1123] MS: 532

[1124]118at.2(R)-Amino-N-{1(S)-[(4-amino-thieno[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-2-naphthalen-1-yl-ethyl-4-phenyl-butyramide

[1125] MS: 538

Example 119

[1126] The general procedure described in example 116 was repeated.After cleavage of the product from the resin, the organic phases werecombined. Half of the solution was used to isolate the N-Boc-protectedcompounds, the other half was used to deprotect the compounds. Thepresence of the desired compounds was detected by mass spectrum, andtheir purities were determined by TLC. The solvents were removed invacuo to give the following pairs of compounds:

[1127] 119a1.3(R)-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-azetidine-1-carbonyl}-3,4-dihydro-1H-isoquinoline-2-carboxylicacid tert-butyl ester

[1128] MS: 516

[1129] and

[1130] 119a2.1-(1,2,3,4-Tetrahydro-isoquinoline-3(R)-carbonyl)-azetidine-2(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide

[1131] MS: 416

[1132] 119b1.3(R)-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidine-1-carbonyl}-3,4-dihydro-1H-isoquinoline-2-carboxylicacid tert-butyl ester

[1133] MS: 530

[1134] and

[1135] 119b2.1-(1,2,3,4-Tetrahydro-isoquinoline-3(R)-carbonyl)-pyrrolidine-2(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide

[1136] MS: 430

[1137] 119c1.[2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-(1H-indol-3-ylmethyl)-2-oxo-ethyl]-carbamicacid tert-butyl ester

[1138] MS: 557

[1139] and

[1140] 119c2.1-[2(R)-Amino-3-(1H-indol-3-yl)-propionyl]-pyrrolidine-2(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide

[1141] MS: 457

[1142] 119d1.[2-{2(S)-[(4Amino-thieno[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-(1H-indol-3-ylmethyl)-2-oxo-ethyl]-carbamicacid tert-butyl ester

[1143] MS: 563

[1144] and

[1145] 119d2.1-[2(R)-Amino-3-(1H-indol-3-yl)-propionyl]-pyrrolidine-2(S)-carboxylicacid (4-amino-thieno[3,2-c]pyridin-2-ylmethyl)-amide

[1146] MS: 463

[1147] 119e1.[2-{2(S)-[(4Amino-thieno[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-(1-formyl-1H-indol-3-ylmethyl)-2-oxo-ethyl]-carbamicacid tert-butyl ester

[1148] MS: 591

[1149] and

[1150] 119e2.1-[2(R)-Amino-3-(1formyl-1H-indol-3-yl)-propionyl]-pyrrolidine-2(S)-carboxylicacid (4-amino-thieno[3,2-c]pyridin-2-ylmethyl)-amide

[1151] 119f1.(2-{2(S)-[(4-Amino-thieno[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-oxo-1(R)-phenyl-ethyl)-carbamicacid tert-butyl ester

[1152] MS: 510

[1153] and

[1154] 119f2. 1-((R)-Amino-phenyl-acetyl)-pyrrolidine-2(S)-carboxylicacid (4-amino-thieno[3,2-c]pyridin-2-ylmethyl)-amide

[1155] MS: 410

[1156] 119g1.[2-{2(S)-[(4-Amino-thieno[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-(4-methoxy-benzyl)-2-oxo-ethyl]-carbamicacid tert-butyl ester

[1157] MS: 554

[1158] and

[1159] 119g2.1-[2(R)-Amino-3-(4-methoxy-phenyl)-propionyl]-pyrrolidine-2(S)-carboxylicacid (4-amino-thieno[3,2-c]pyridin-2-ylmethyl)-amide

[1160] MS: 454

[1161] 119h1.[2-{2(S)-[(4-Amino-furo[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-(4-methoxy-benzyl)-2-oxo-ethyl]-carbamic acid tert-butylester

[1162] MS: 538

[1163] and

[1164] 119h2.1-[2(R)-Amino-3-(4-methoxy-phenyl)-propionyl]-pyrrolidine-2(S)-carbolicacid (4-amino-furo[3,2-c]pyridin-2-ylmethyl)-amide

[1165] MS: 438

[1166] 119i1.[2-{2(S)-[(4-Amino-thieno[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-(4-fluoro-benzyl)-2-oxo-ethyl]-carbamicacid tert-butyl ester

[1167] MS: 542

[1168] and

[1169] 119i2.1-[2(R)-Amino-3-(4-fluoro-phenyl)-propionyl]-pyrrolidine-2(S)-carboxylicacid (4-amino-thieno[3,2-c]pyridin-2-ylmethyl)-amide

[1170] MS: 442

[1171] 119j1.[2-{2(S)-[(4-Amino-thieno[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-(4-chloro-benzyl)-2-oxo-ethyl]-carbamicacid tert-butyl ester

[1172] MS: 559

[1173] and

[1174] 119j2.1-[2(R)-Amino-3-(4-chloro-phenyl)-propionyl]-pyrrolidine-2(S)-carboxylicacid (4-amino-thieno[3,2-c]pyridin-2-ylmethyl)-amide

[1175] MS: 459

[1176] 119k1.(3-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-naphthalen-1-ylmethyl-2-oxo-ethyl)-carbamicacid tert-butyl ester

[1177] MS: 568

[1178] and

[1179] 119k2.1-(2(R)-Amino-3-naphthalen-1-yl-2-oxo-propyl)-pyrrolidine-2(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide

[1180] MS: 482

[1181] 119l1.(1(R)-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-pyrrolidine-1-carbonyl}-3-phenyl-propyl)-carbamicacid tert-butyl ester

[1182] MS: 532

[1183] and

[1184] 119l2.1-(2(R)-Amino-4-phenyl-butyryl)-pyrrolidine-2(S)-carboxylic acid(1-amino-isoquinolin-6-ylmethyl)-amide

[1185] MS: 432

[1186] 119m1.(2-{2(S)-[(1-Amino-isoquinolin-7-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-benzyl-2-oxo-ethyl)-carbamicacid tert-butyl ester

[1187] MS: 518

[1188] and

[1189] 119m2.1-(2(R)-Amino-3-phenyl-propionyl)-pyrrolidine-2(S)-carboxylic acid(1-amino-isoquinolin-7-ylmethyl)-amide

[1190] MS: 418

[1191] 119n1.[2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-azetidin-1-yl}-1(R)-(4-methoxy-benzyl)-2-oxo-ethyl-carbamicacid tert-butyl ester

[1192] MS: 534

[1193] and

[1194] 119n2.1-[2(R)-Amino-3-(4-methoxy-phenyl)-propionyl]-azetidine-2(S)-carboxylicacid (1-amino-isoquinolin-6-ylmethyl)-amide

[1195] MS: 434

[1196] 119o1.[2-{2(S)-[(1-Amino-isoquinolin-7-ylmethyl)-carbamoyl]-azetidin-1-yl}-1(R)-(4-methoxy-benzyl)-2-oxo-ethyl-carbamicacid tert-butyl ester

[1197] MS: 534

[1198] and

[1199] 119o2.1-[2(R)-Amino-3-(4-methoxy-phenyl)-propionyl]-azetidine-2(S)-carboxylicacid (1-amino-isoquinolin-7-ylmethyl)-amide

[1200] MS: 434

[1201] 119p1.[2-{2(S)-[(1-Amino-isoquinolin-7-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-1(R)-(4-methoxy-benzyl)-2-oxo-ethyl]-carbamicacid tert-butyl ester

[1202] MS: 548

[1203] and

[1204] 119p2.1-[2(R)-Amino-3-(4-methoxy-phenyl)-propionyl]-pyrrolidine-2(S)-carboxylicacid (1-amino-isoquinolin-7-ylmethyl)-amide

[1205] MS: 448

[1206] 119q1.(2-{2(S)-[(1-Amino-isoquinolin-6-ylmethyl)-carbamoyl]-azetidin-1-yl}-2-oxo-1(R)-phenyl-ethyl)-carbamicacid tert-butyl ester

[1207] MS: 490

[1208] and

[1209] 119q2. 1-((R)-Amino-phenyl-acetyl)-azetidine-2(S)-carboxylic acid(1-amino-isoquinolin-6-ylmethyl)-amide

[1210] MS: 390

[1211] 119r1.(2-{(2(S)-[(1-Amino-isoquinolin-7-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-oxo-1(R)-phenyl-ethyl)-carbamicacid tert-butyl ester

[1212] MS: 504

[1213] and

[1214] 119r2. 1-((R)-Amino-phenyl-acetyl)-pyrrolidine-2(S)-carboxylicacid (1-amino-isoquinolin-7-ylmethyl)-amide

[1215] MS: 404

[1216] 119s1.(2-{2(S)-[(4-Amino-thieno[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-oxo-ethyl)-phenyl-carbamicacid tert-butyl ester

[1217] MS: 510

[1218] and

[1219] 119s2. 1-Phenylaminoacetyl-pyrrolidine-2(S)-carboxylic acid(4-amino-thieno[3,2-c]pyridin-2-ylmethyl)-amide

[1220] MS: 410

[1221] 119t1.(2-{2(S)-[(4-Amino-thieno[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-oxo-ethyl)-benzyl-carbamicacid tert-butyl ester

[1222] MS: 524

[1223] and

[1224] 119t2. 1-Benzylaminoacetyl-pyrrolidine-2(S)-carboxylic acid(4-amino-thieno[3,2-c]pyridin-2-ylmethyl)-amide

[1225] MS: 424

[1226] 119u1.(2-{2(S)-[(4-Amino-thieno[3,2-c]pyridin-2-ylmethyl)-carbamoyl]-pyrrolidin-1-yl}-2-oxo-ethyl)-phenylethyl-carbamicacid tert-butyl ester

[1227] MS: 538

[1228] and

[1229] 119u2. 1-Phenethylaminoacetyl-pyrrolidine-2(S)-carboxylic acid(4-amino-thieno[3,2-c]pyridin-2-ylmethyl)-amide

[1230] MS: 438

Example 120

[1231] The following compounds were prepared using the proceduresdescribed in example 73.

[1232] 120a. N-(1-amino-isoquinolin-6-ylmethyl)1-((2-(hydroxycarbonylmethylamino)-2-(cyclooctylmethyl)-acetyl)-prolinamidehydrochloride

[1233] Using 306 mg of1-((2-((butyloxycarbonyl)(butyloxycarbonylmethyl)amino)-2-(cyclooctylmethyl)-acetyl)-proline(prepared according to WO 97/31939 example 7) and 104 mg of1-amino-6-(aminomethyl)isoquinoline gave 62 mg of the title compound. MSESI+: 510 (M+H).

[1234] 120b. N-(1-amino-isoquinolin-6-ylmethyl)1-(2(R)-(hydroxycarbonylmethylamino)-1-oxo-hexyl)-prolinamidetrifluoroacetate

[1235] The use of 420 mg of1-((2-((butyloxycarbonyl)(butyloxycarbonylmethyl)amino)-(2(R)-(hydroxycarbonylmethylamino)-1-oxo-hexyl)-proline( prepared according to WO 97/31939 example 48) and 165 mg of1-amino-6-(aminomethyl)isoquinoline gave 330 mg of the title compound.MS ESI+: 442 (M+H).

[1236] 120c.N-(1-amino-isoquinolin-6-ylmethyl)-4-cis-ethyl-1-((2-(hydroxycarbonylmethylamino)-2(R)-(cyclohexylmethyl)-acetyl)-prolinamidetrifluoroacetate

[1237] Starting with 345 mg of1-((2-((butyloxycarbonyl)(butyloxycarbonylmethyl)amino)-2-(cyclohexylmethyl)-acetyl)-4-cis-ethyl-proline(prepared according to WO 97/31939 example 52) and 122 mg of1-amino-6-(aminomethyl)isoquinoline gave 412 mg of the title compound.

[1238] MS ESI+: 510(M+H).

[1239] 120d. N-(1-amino-isoquinolin-6-ylmethyl)1-cyclopentyl-1-(2-((2-propyl)oxycarbonyl-methylamino)-2(R)-cyclohexylmethyl-acetyl)glycinamidehydrochloride

[1240] The use of 397 mg of1-cyclopentyl-1-(2-((2-propyl)oxycarbonylmethylamino)-2(R)-cyclohexylmethyl-acetyl)glycineand 138 mg of 1-amino-6-(aminomethyl)isoquinoline gave 301 mg of thetitle product.

[1241] MS ESI+: 552(M+H).

Example 121 N-(1-amino-isoquinolin-6-ylmethyl)1-(2-((2-propyl)oxycarbonylmethylamino)-2(R)-cyclohexylmethyl-acetyl)-prolinamide

[1242] To a stirred solution of 96 mg ofN-(1-amino-isoquinolin-6-ylmethyl)1-(2-hydroxycarbonylmethylamino-2(R)-cyclohexylmethyl-acetyl)-prolinamidein 6 mL of 2-propanol was added 0.03 mL of thionyl chloride and thereaction mixture was heated at refux temperature for two days. Afterthis period the reaction mixture was concentrated, ethyl acetate added,washed with aqueous 5% sodium hydrogencarbonate and brine, dried oversodium sulphate and concentrated. The residue was dissolved in at-butanol/water 1/1 (v/v) mixture lyophilisation yielded 56 mg of thetitle compound. MS ESI+: 524 (M+H)

Example 122

[1243] The following compounds were prepared using the proceduresdescribed in example 121.

[1244] 122a.N-(1-amino-isoquinolin-6-ylmethyl)1-(2-((1-butyl)oxycarbonylmethylamino)-2(R)-cyclohexylmethyl-acetyl)-prolinamide

[1245] Yield: 47%, MS ESI+: 538 (M+H).

[1246] 122b.N-(1-amino-isoquinolin-6-ylmethyl)1-(2-((2-methoxyethyl)oxycarbonylmethylamino)-2(R)-cyclohexylmethyl-acetyl)-prolinamide

[1247] Yield: 70%, MS ESI+: 540 (M+H).

[1248] 122c. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-(benzyloxycarbonylmethylamino)-2(R)-cyclohexylmethyl-acetyl)-prolinamide

[1249] Yield: 32%, MS ESI+: 572 (M+H).

[1250] 122d.N-(1-amino-isoquinolin-6-ylmethyl)1-(2(R)-(propyyoxycarbonylmethylamino)-1-oxo-hexyl)-prolinamidehydrochloride

[1251] Hydrochloric acid was added to the t-butanol/water solution ofthe free base and lyophilistation afforded the title compound: Yield:73%, MS ESI+: 484(M+H).

[1252] 122e.N-(1-amino-isoquinolin-6-ylmethyl)-4-cis-ethyl-1-((2-(propyloxycarbonylmethylamino)-2(R)-(cyclohexylmethyl)-acetyl)-prolinamide

[1253] Yield. 51%, MS ESI+: 552 (M+H).

[1254] 122f.N-(1-amino-isoquinolin-6-ylmethyl)-1-((2-(propyloxycarbonylmethylamino)-2(R)-((4-methoxyphenyl)methyl)-acetyl)-prolinamide

[1255] Yield: 90%, MS ESI+: 548 (M+H).

Example 123N-(1-amino-isoquinolin-6-ylmethyl)1-(2-((morpholine-4-yl)carbonylmethylamino)-2(R)-cyclohexylmethyl-acetyl)-prolinamidetrifluoroacetate

[1256] 123a. N-Boc-N-((morpholin-4-yl)carbonylmethyl)-D-Cha-OH

[1257] Using the procedures described in example 73 2.0 g of H-D-Cha-OMe. HCl and 2.07 g of 4-(bromoacetyl)morpholine (J. Med. Chem. 35, 1685(1992)) gave 1.77 g of the title compound. Rf=0.22 in ethylacetate/heptanes=6/4 (v/v) on silica.

[1258] 123b. N-Boc-N-((morpholin-4-yl)carbonylmethyl)-D-Cha-Pro-OH

[1259] To a stirred solution of 0.86 g ofN-Boc-N-((morpholin-4-yl)carbonylmethyl)-D-Cha-OH in 8 mL ofN,N-dimethylformamide at 0° C. were successively added 0.45 g of1-hydroxy benzotriazole (HOBT) and 0.51 g of dicyclohexyl carbodimide(DCC). After 30 minutes a mixture of 0.58 g of proline benzylesterhydrochloride and 0.13 mL of triethyl amine in 8 nL ofN,N-dimethylformamide was added. The reaction mixture was allowed tocome to room temperature and an additional 0.05 mL of triethylamine wasadded. After 16 hours 0.36 g of DCC and 0.20 mL of triethylamine wereadded and stirred for 3 days at room temperature. . The mixture wascooled to −20° C. and dicyclohexylurea was removed by filtration. Thefiltrate was evaporated to dryness The residue was dissolved in ethylacetate and washed successively with 5% sodium hydrogencarbonate, water,2% citric acid and brine, dried over sodium sulphate and concentrated invacuo. The residue was chromatographed on silica gel eluting withdichloromethane/methanol: 3/1 v/v as eluent to yield 1.02 g ofN-Boc-N-((morpholin-4-yl)carbonylmethyl)-D-Cha-Pro-OBzl This benzylester was dissolved in 10 mL of methanol, 125 mg of 10% palladium oncharcoal was added and the mixture was hydrogenated at atmosphericpressure at room temperature for 2 hours. The palladium catalyst wasremoved by filtration and the solvent removed by evaporation at reducedpressure to yield 0.86 g of the title compound.

[1260] MS ESI: 494 (M−H)⁻.

[1261] 123c.N-(1-amino-isoquinolin-6-ylmethyl)1-(2-((morpholin-4-yl)carbonylmethylamino)-2(R)-cyclohexylmethyl-acetyl)-prolinamidetrifluoroacetate

[1262] This compound was prepared using the procedures described inexample 73. A TBTU-coupling of 150 mg ofN-Boc-N-((morpholin-4-yl)carbonylmethyl)-D-Cha-Pro-OH and 58 mg1-amino-6-(aminomethyl)isoquinoline and deprotection usingtrifluoroacetic acid yielded 186 mg of the title compound.

[1263] MS ESI⁺: 551 (M+H).

Example 124

[1264] The following compounds were prepared using the proceduresdescribed in example 1 k.

[1265] 124a. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-((dimethylamino)carbonylmethylamino)-2(R)-cyclohexylmethyl-acetyl)-prolinamidehydrochloride

[1266] DCC/HOBt coupling of 193 mg of N-(1-amino-isoquinolin-6-ylmethyl)1-(2-hydroxycarbonylmethylamino-2(R)-cyclohexylmethyl-acetyl)-prolinamideand 65 mg of dimethylamine hydrochloride yielded 40 mg of the titlecompound. MS ESI⁺: 509 (M+H).

[1267] 124b. N-(1-amino-isoquinolin-6-ylmethyl)1-(2-(azetidin-1-ylcarbonylmethylamino)-2(R)-cyclohexylmethyl-acetyl)-prolinamidehydrochloride

[1268] DCC/HOBt coupling of 193 mg of N-(1-amino-isoquinolin-6-ylmethyl)1-(2-hydroxycarbonylmethylamino-2(R)-cyclohexylmethyl-acetyl)-prolinamideand 42 mg of azetidine hydrochloride yielded 40 mg of the titlecompound. MS ESI⁺: 521 (M+H).

Example 125(4-(4-((1-aminoisoquinolin-7-yl)methylaminocarbonyl)benzoyl)-piperazin-1-yl)acetate

[1269] Bromoacetate was coupled to the resin via an ester linkage.Piperazine was introduced followed by a peptide coupling withterephthalic acid. Then 1-amino-7-(aminomethyl)isoquinoline was coupledand finally the product was cleaved off and lyophilisation gave thetitle compound.

[1270] MS (IonSpray): 447.5

Example 126

[1271] The following compounds were prepared using the proceduredescribed in example 125.

[1272] 126a.(4-(3-((1-aminoisoquinolin-7-yl)methylaminocarbonyl)benzoyl)-piperazin-1-yl)acetate,

[1273] MS (IonSpray): 447.5

[1274] 126b.(4-(2-((1-aminoisoquinolin-7-yl)methylaminocarbonyl)cyclohexyicarbonyl)-piperazin-1-yl)acetate,MS (IonSpray): 453.6

[1275] 126c.(4-(4-((1-aminoisoquinolin-7-yl)methylamino)1,4-dioxo-2-buten-1-yl)-piperazin-1-yl)acetate,MS (IonSpray): 397.5

[1276] 126d.((3-(4-((1-aminoisoquinolin-7-yl)methylaminocarbonyl)benzoyl)(methyl)amino)propyl)(methyl)amino)acetate, MS (IonSpray): 449.5

[1277] Other compounds of the invention which may be prepared followingthe procedures as set out in the specification are shown in Tables Ia,Ib, Ib, Ic, Id, Ie, If, Ig, Ig, Ih, Ij Ik. and Il TABLE Ia

X = CH═CH, O, S

[1278] TABLE Ib

X = CH═CH, O, S

[1279] TABLE Ic

R = OCH(CH₃)₂ N(CH₃)₂ NH(C(CH₃)₃)

p = 2, 3, 4

[1280] TABLE Id

R¹SO₂:

[1281] TABLE Ie

R¹SO₂:

[1282] TABLE If

R¹SO₂:

[1283] TABLE Ig

R¹SO₂:

[1284] TABLE Ih

R¹SO₂:

[1285] TABLE Ij

X═CH═CH, O, S

[1286] TABLE Ik

E:

[1287] TABLE Il

J-D:

[1288] Also the following compounds may be prepared according theprocedures as previously described:

[1289] The biological activities of the compounds of the presentinvention were determined by the following test methods.

I. Anti-thrombin assay

[1290] Thrombin (Factor IIa) is a factor in the coagulation cascade. Theanti-thrombin activity of compounds of the present invention wasassessed by measuring spectrophotometrically the rate of hydrolysis ofthe chromogenic substrate s-2238 exterted by thrombin. This assay foranti-thrombin activity in a buffer system was used to assess theIC₅₀-value of a test compound. Test medium:Tromethamine-NaCl-polyethylene glycol 6000 (TNP) buffer Referencecompound: I2581 (Kabi) Vehicle: TNP buffer Solubilisation can beassisted with dimethylsulphoxide, methanol, ethanol, acetonitrile ortert.-butyl alcohol which are without adverse effects in concentrationsup to 2.5% in the final reaction mixture. Technique Reagents* 1.Tromethamine-NaCl (TN) buffer   Composition of the buffer: Tromethamine(Tris) 6.057 g (50 mmol) NaCl 5.844 g (100 mmol) Water to 1 l   The pHof the solution is adjusted to 7.4 at 37° C. with HCl (10   mmol · l⁻¹).2. TNP buffer   Polyethylene glycol 6000 is dissolved in TN buffer togive a   concentration of 3 g · l⁻¹ 3. S-2238 solution   One vial S-2238(25 mg; Kabi Diagnostica, Sweden) is dissolved in 20   ml TN buffer togive a concentration of 1.25 mg · ml⁻¹ (2 mmol · l⁻¹). 4. Thrombinsolution   Human thrombin (16 000 nKat · vial⁻¹; Centraal Laboratoriumvoor   Bloedtransfusie, Amsterdam, The Netherlands) is dissolved in TNP  buffer to give a stock solution of 835 nKat · ml⁻¹.   Immediatelybefore use this solution is diluted with TNP buffer to give   aconcentration of 3.34 nKat · ml⁻¹.

[1291] Preparation of test and reference compound solutions

[1292] The test and reference compounds are dissolved in Milli-Q waterto give stock concentrations of 10⁻² mol·1⁻¹. Each concentration isstepwise diluted with the vehicle to give concentrations of 10⁻³, 10⁻⁴and 10⁻⁵ mol·1⁻¹. The dilutions, including the stock solution, are usedin the assay (final concentrations in the reaction mixture: 3·10⁻³;10⁻³; 3·10⁻⁴; 10⁻⁴; 3·10⁻⁵; 10⁻⁵; 3·10⁻⁶ and 10⁻⁶ mol·1⁻¹, respectively.

[1293] Procedure

[1294] At room temperature 0.075 ml and 0.025 ml test compound orreference compound solutions or vehicle are alternately pipetted intothe wells of a microtiter plate and these solutions are diluted with0.115 ml and 0.0165 ml TNP buffer, respectively. An aliquot; of 0.030 mlS-2238 solution is added to each well and the plate is pre-heated andpre-incubated with shaking in an incubator (Amersham) for 10 min. at 37°C. Following pre-incubation the hydrolysis of S-2238 is started byaddition of 0.030 ml thrombin solution to each well. The plate isincubated (with shaking for 30 s) at 37° C. Starting after 1 min ofincubation, the absorbance of each sample at 405 nm is measured every2,min. for a period of 90 min. using a kinetic microtiter plate reader(Twinreader plus, Flow Laboratories).

[1295] All data are collected in an IBM personal computer usingLOTUS-MEASURE. For each compound concentration (expressed in mol·1⁻¹reaction mixture) and for the blank the absorbance is plotted versus thereaction time in min.

[1296] Evaluation of responses:

[1297] For each final concentration the maximum absorbance wascalculated from the assay plot. The IC₅₀-value (final concentration,expressed in μmol·1⁻¹, causing 50% inhibition of the maximum absorbanceof the blank) was calculated using the logit transformation analysisaccording to Hafner et al. (Arzneim.-Forsch./Drug Res. 1977; 27(II):1871-3). Antithrombin activity: Example IC₅₀ (μmol · l⁻¹) 5 28 7 6.8 80.082 10 1.1 18 0.53 30 35 32 1.44

II. Anti-factor Xa assay

[1298] Activated Factor X (Xa) is a factor in the coagulation cascade.The anti-Xa activity of compounds of the present invention was assessedby measuring spectrophotometrically the rate of hydrolysis of thechromogenic substrate s-2222 exterted by Xa. This assay for anti-Xaactivity in a buffer system was used to assess the IC₅₀-value of thetest compound.

[1299] In general the followed procedure and test conditions wereanalogous to those of the anti-thrombin assay as described above.Differences are indicated below. Reference compound: benzamidineVehicle: TNP buffer. Solubilisation can be assisted withdimethylsulphoxide, methanol, ethanol, acetonitrile or tert.-butylalcohol which are without adverse effects in concentrations up to 1%(for DMSO) and 2.5% (for the other solvents) in the final reactionmixture. Technique Reagents* 3. S-2222 solution   One vial S-2222 (15mg; Kabi Diagnostica, Sweden) is dissolved in 10   ml water to give aconcentration of 1.5 mg · ml⁻¹ (2 mmol · l⁻¹). 4. Xa solution   BovineFactor Xa Human (71 nKat · vial⁻¹; Kabi Diagnostica) is   dissolved in10 ml TNP buffer and then further diluted with 30 ml   TNP buffer togive a concentration of 1.77 nKat · ml⁻¹. The dilution has   to befreshly prepared. Procedure Instead of the S-2238 solution (inanti-thrombin assay), the above S-2222 solution is added to each well inthis assay.

[1300] Anti-factor Xa activity Example IC₅₀ (μmol · l⁻¹) 1 48 3 29 14 4124 14 26 14

III. Caco-2 permeability

[1301] Caco-2 cells were obtained from the American Type CultureCollection and were used in passage nrs 25 to 35. Cells were maintainedat 37° C. in 75 cm² culture flasks (Nunc) in Dulbecco's Modified Eagle'sMedium (DNEM) supplemented with 1% of non-essential amino acid solution(100×), 10% of heat-inactivated fetal bovine serum, penicillin (100U/ml) and streptomycin (100 μg/ml) in an atmosphere of 95% of air and 5%of CO₂. The pH of the medium was 7.4. For experiments cells were grownon Transwell-COL™ collagen treated cell culture filter inserts (Costar,Cambridge, Mass., USA) with a surface area of 0.33 cm² The seedingdensity was 6.3 ×10⁵ cells/cm². Culture medium was added on each side ofthe filter. The growth of the cells and degree of confluency was checkedevery 2-3 days microscopically. Confluent monolayers were used on day23-24 after seeding, at which time the transepithelial electricalresistance of the monolayers was at a stable value of approximately 260Ω.cm². Permeability experiments were performed with compoundconcentrations of 0.1. mM or 1 mM in the apical compartment in Hanks'Balanced Salt Solution with or without 0.5% (w/v) of bovine serumalbumin. At 1, 2 and 3 hours after addition of compound to the cells thefilters were placed into fresh acceptor compartments. In this way threeone-hour samples were obtained. Compound concentration was determined insamples from acceptor compartments and donor compartment by HPLC or bymeasuring anti-thrombin activity. Permeability experiments of eachcompound were performed with 4 different filters. Results shown in theTable are means of the permeability between 2 and 3 hours after thestart of the experiment of at least 2 filters.

[1302] In the following Table the apparent Caco-2 permeability (P_(app))is given in nm/s for compounds of the present invention compared to theprior art compounds NAPAP and “Pefa 1286”*. Compound Caco-2 P_(app)(nm/s) NAPAP 4 “Pefa 1286”* 1.1 ex.2  19 ex.4  25 ex.5  37 ex.11 148ex.13 118 ex.14 10 ex.18 121

Conclusion:

[1303] The apparent permeability of compounds of the present inventionis significantly higher than that of the prior art compounds NAPAP andPefa 1286. Compounds of the present invention will therefore beconsiderably better absorbed in the gastrointestinal tract.

We claim:
 1. A serine protease inhibitor of formula (XL),

wherein (R″′)¹ and (R″′)² are independently H, lower alkyl, aryl,heteroaryl, cycloalkyl or lower alkyl substituted by one or moresubstituents selected from CONH₂, COO— (lower alkyl), aryl, asheteroaryl and cycloalkyl; or (R″′)² is lower alkanoyl; (R″′)³ is H,COOH, CONH₂, COO— (lower alkyl), CONH— (lower alkyl) or CON(loweralkyl)₂; (R″′)⁴, (R″′)⁵ and (R″′) are independently H, lower alkyl,aryl, aralkyl or cycloalkyl; or (R″′)⁴ and/or one of (R″′)⁵ and (R″′)⁶is heteroaryl or lower alkyl substituted with OH, SO₂H, SO₃H, guanidino,aryl-(lower alkoxy), lower alkoxy or lower alkylthio; or (R″′)² togetherwith (R″′)⁴ forms a tri- or tetramethylene group, in which (a) amethylene group may be replaced by S, SO or SO₂ or may be substitutedwith OH, lower alkyl, lower alkenyl or carboxy-(lower alkyl) or (b) oneof the methylene groups may be substituted with lower alkenyl and theother with (lower alkyl) —COOH; and at least one of (R″′)¹, (R″′)²,(R″′)⁴, (R″′)⁵ and (R″′)⁶ is the group of formula Q, wherein Q isrepresented by formula

wherein the substructure

is a structure selected from

wherein X is C or S; X′ being independently CH or N; and m is 0, 1, 2 or3; wherein the group Q is bound through an oxygen atom or anunsubstituted or substituted nitrogen or carbon atom.
 2. The serineprotease inhibitor of claim 1, wherein Q has the formula

m is 0, 1, 2 or
 3. 3. A pharmaceutical composition, comprising: theserine protease inhibitor of claim 1 and a pharmaceutically acceptableauxiliary.
 4. A method of treating or preventing thrombosis orthrombosis-related diseases in a patient in need thereof, comprising:administering to the patient an effective amount of a compound accordingto claim
 1. 5. A process for preparing a pharmaceutical composition,comprising: mixing together a compound of claim 1 with pharmaceuticalacceptable auxiliaries.